Protein Engineering _ Siti Julaiha
-
Author
siti-julaiha-gruebner -
Category
Documents
-
view
227 -
download
0
Embed Size (px)
Transcript of Protein Engineering _ Siti Julaiha
-
7/23/2019 Protein Engineering _ Siti Julaiha
1/98
Jck)ici`hzh lsked sk}sl lcuhihhk ld}h mhk zhpduhk lcskdlhk ihudkj ihudkj
dkmdqdms+
Lc}dlh jck `clcpfh mckjhk `hdl/ }s`sb ld}h `cplci`hkj mhk icidodld askjud thkj
`hdl+ ]h~d `doh uc`shb jck }skjjho ) `hblhk ucjick lcedo mhpd uc`shb jck }skjjho ickjhohid
lclhehshk/ mh~h} `cpcacl mckjhk hmhkth eheh} mhk ~ckthld}/ `hblhk lcih}dhk+
Jck hmhohb us`skd} ~clcpfh mhpd MKH+
MKH hmhohb `osc ~pdk} mhpd uc}dh~ dkmdqshodu gpjhkdui+ Icps~hlhk mh}h`huc dkagpihud
ldidh `cuhp thkj ici`hzh uc} dku}pslud ockjlh~ sk}sl ici`sh} ucish ~pg}cdk thkj hlhk
md`s}sblhk }s`sb+ Uc}dh~ jck `cpdud ucphkjlhdhk dku}pslud }cp}ck}s/ `dhuhkth ickjlgmclhk
~pg}cdk }cp}ck}s+ Hjhp uco mh~h} ici`sh} ~pg}cdk/ dkagpihud mhpd jck mduhodk/ `hudu mckjhk
`hudu/ mhpd MKH lc mhohi hosp `hps mhpd icuuckjcp PKH "iPKH$+ Lcismdhk iPKH `cpfhohkhk
lcoshp kslocsu icksfs lc ud}g~ohuih/ mhk ~hmh `hjdhk uco thkj mduc`s} gpjhkco pd`gugi/
iPKH ickjhphblhk ~cphld}hk huhi hidkg thkj }cpod~h} ickfhmd igoclso ~pg}cdk+
MKH bhmdp uc`hjhd msh ~huhkj phk}hd ~hkfhkj/ icodkjlhp mhohi `ck}sl bcodlu
jhkmh+ Uc}dh~ sk}hd }cpmdpd mhpd fs}hhk `ogl `hkjskhk ldidh thkj mduc`s} `huc+ Ucick}hph
bhkth hmh ci~h} mhuhp ldidh thkj `cp`cmh mhohi MKH thd}s Hmckdk/ ]didk/ Ud}gudk/ mhk
Jshkdk/ sps}hk `huc)`huc thkj }cpfhmd ickck}slhk dkagpihud thkj }cpucmdh+
MKH `cphmh ~hmh dk}d/ h}hs kslocsu/ ihudkj)ihudkj mhpd }pdodskhk uco }s`sb+ Uc}dh~ uco
ihksudh "mckjhk ~ckjceshodhk uco mhphb icphb/ thkj }dmhl icidodld dk}d$ `cpdud MKH thkj
uhih+ Uc}dh~ uco icidodld =1 igoclso MKH sk}hd jhkmh+ Uc}dh~ igoclso }cpmdpd mhpd 3>)
03>>>>>>> hudu }cpudi~hk ~hmh+ lpgigugi+
MKH uc}dh~ lpgigugi ickjhkmskj `hkthl jck mhk ici`ck}hkj oshu/ }cpihusl kgk
lgmdkj MKH thkj askjudkth `cosi mdlc}hbsd+
H~h d}s ^pg}cdk7
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 8
@DG]CLKGOGJD
PCLHTHUH ^PG]CDK
Ud}d Fsohdbh ' >4>134:=0>
^hueh Uhpfhkh]clkgogjd @dgicmdu
Skdqcpud}hu Dkmgkcudh
^hueh Uhpfhkh ]clkgogjd @dgicmduSkdqcpud}hu Dkmgkcudh
\Icd 0>8>Y
-
7/23/2019 Protein Engineering _ Siti Julaiha
2/98
^pg}cdk hmhohb ususkhk phk}hd ~godicp thkj }cp`ck}sl mhpd udk}cuh huhi hidkg huhi hidkg
mhohi phkjlhdhk lgi~oclu `cp`hjhd fckdu dlh}hk mhk u}psl}sp ucbdkjjh }cp`ck}sl fckdu fckdu
~pg}cdk thkj `cp`cmh uh}s uhih ohdk+
Mhohi }s`sb ihksudh/ ~pg}cdk }cp`ck}sl mhpd mhpd udk}cuh thkj }cpfhmd mhohi uco mdisohd mhpd
udk}cuh phk}hd jhkmh MKH thkj }cpmh~h} mhohi kslocsu uco thkj ickjhohid ~pgucu udk}cuh}phkulpd~ud ickfhmd phk}hd }skjjho PKH mhk mhpd phk}hd PKH }cpuc`s} ickjhohid ~pgucu
}phkulpd~ud mckjhk hmhkth pd`gugi ucbdkjjh }cp`ck}sl ~pgucu ~ci`ck}slhk phk}hd jhkmh
mckjhk ~pgucu ~ci`ck}slhk agomdkj h}hs od~h}hk od~h}hk phk}hd ickfhmd phk}hd lgi~oclu mhohi
u}psl}sp thkj `cp`cmh+
PCLHTHUH ^PG]CDK
U}psl}sp ~pg}cdk thkj `cp`cmh icktc`h`lhk }cpfhmdkth ~cp`cmhhk askjud fsjh ~hmh ~pg}cdk
~pg}cdk thkj }cp`ck}sl mhohi uco+
Lhphl}cpdu}dl huhi hidkg uhkjh} md~cposlhk lc}dlh pclhthuh ~pg}cdk mdjskhlhk sk}sl icisohd
ush}s ~pgtcl+ @dhuhkth ~pgtcl thkj `cp}sfshk icohlslhk ~cps`hbhk ~hmh ~pg}cdk thkj }cohb
cludu h}hs hmh/ icisohd ohkjlhb hzhokth mckjhk ~ckck}shk huhi hidkg thkj mdjskhlhk
sk}sl icked~}hlhk is}hud+ Bho dkd uhkjh} ~ck}dkj lhpckh lgk}pd`sud huhi hidkg }cpbhmh~
askjud ~pg}cdk icps`hb udah} mhk lhphl}cpdu}dl ~pg}cdk/ ucbdkjjh ~ckjc}hbshk h}hs hkhoduh
}cpbhmh~ lhphl}cpdu}dl huhi hidkg bhpsu }cohb md~dldplhk+ Lc}dlc icpclhthuh ~pg}cdk sk}sl
askjud mhk lhphl}cpdudl kgqco'`hps/ bho thkj }dmhl sisi khisk ickhpdl sk}sl mdohlslhk
hmhohb ickjh}sp `c`cph~h is}hk ickjjskhlhk huhi hidkg thkj `cpqhpdhud mhk fsjh
icpclhthuh is}hud ucehph iso}d~oc ~hmh uh}s ~pg}cdk+
Pclhthuh ~pg}cdk hmhohb h~oodlhud ehi~sphk ickhpdl mhpd ~ckjc}hbshk 6
) @dgogjd Igoclsocp
) Hkhoduh U}psl}sp Ldidh pg}cdk
) Ih}cih}dlh
) Clgkgid
) Lgi~s}hud h}hs @dg Dkagpih}dlh
) @dgldidh
sk}sl ici~pgucu ~ckjci`hkjhk ~pg}cdk ickfhmd ~pgmsl thkj `cpjskh mhk `cpbhpjh+
Icps~hlhk mdud~odk dois thkj mdlh}cjgpdlhk ismh sudhkth/ mckjhk ~ckcod}dhk ~ckcod}dhk thkj
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 0
-
7/23/2019 Protein Engineering _ Siti Julaiha
3/98
uhh} dkd mdohlslhk sk}sl icihbhid u}psl}sp dlh}hk ~pg}cdk/ pclgjkhud ~pg}cdk sk}sl ~pdkud~
~pdkud~ mcuhdk thkj mh~h} md}cph~lhk+
UCFHPHB PCLHTHUH PG]CDK
Pclhthuh ~pg}cdk mdisohd mhpd ~ckcishk ~ckcishk }cpbhmh~ ckvdi mhk askjud
askjudkth/ thd}s ~hmh }hbsk
8255 Hkucoic ^htck! Fchk)Aphkgdu ^cpugv mhpd ^cphkedu/ icohlslhk
Dugohud mhpd lgi~ocl hidohud mhpd jcpidkh}dkj hpoct mhk ickhihlhkkth
mdhu}huc
U}hpeb usjhpu "iho}guc$
8253 F+ Fhlg` @cpvcodku mhpd Uzcmdh
Ickmciguk}phudlhk `hbzhqu}hpeb mh~h} md~cehblhk ucehph cadudck ickjjskhlhkclu}phlud iho}+
Bho thkj mdlckho mckjhk khihLh}hodudu
8251 ]bcgmgp Uebzhkk mhpd Fcpihk
Ickjdugohud ~c~udk/ ckvdi ~cp}hih thkj mducmdhlhk mhpd fhpdkjhk bczhk+
82:1 Zdoodhi Lbkc
Ususkhk }dmhl }cph}sp 'Skgpjhkdvcm acpick acpick mddu}dohblhk ickfhmd Ck vdi thkjmd~c}dl mhpd `hbhuh tskhkd `cphuho mhpd Ck `cphpd dk mhk vtic `cphp}d ohqck'phjd ~hmhtchu}
824: @sebkcp `cpuhsmhph `cphuho mhpd Fcpihk
Ickjclu}phlud ucehph `c`hu uco uco Tchu}
Joseguc c}bhkgo , EG0
]SFSHK PCLHTHUH PG]CDK
) Ickjci`hkjlhk ~pg}cdk sk}sl ucush}s thkj `cpjskh h}hs `cpbhpjh md `dmhkj dkmsu}pd h}hs
lcucbh}hk +
) Ici`ck}sl ~pgmsl hps mhpd ush}hs ehi~sphk ~pg}cdk
) Icps`hb u}psl}sp h}hs udah} ~pg}cdk ucbdkjjh mh~h} mdbhudolhk ~cps`hbhk ~hmh ~pgmsl ~pg}cdk
mckjhk }hpjc} u}psl}sp h}hs udah} thkj mddkjdklhk+
) Icps`hb ~pg}cdk ucbdkjjh mh~h} md}cpfcihblhk sk}sl ici`cpdlhk dkagpihud mhk h~odlhud ohdk+
) Ickjhkhoduh udah} ~pg}cdk thkj hmh sk}sl h~odlhud ~pgmsl ohdk
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 5
-
7/23/2019 Protein Engineering _ Siti Julaiha
4/98
) Ickdkjlh}lhk lcihi~shk mhk cadudckud ~ckjjskhhk ~pgmsl sk}sl ush}s ~pgmsl
LCJDH]HK PCLHTHUH ^PG]CDK
^ckdkjlh}hk lh}hodudu mckjhk us`u}ph} hohid h}hs lgahl}gp
^ckdkjlh}hk lh}hodudu mckjhk us`u}ph} kgkkh}spho h}hs lgahl}gp
^ckdkjlh}hk lh}hodudu md kgkkh}spho ~cohps}
^ckdkjlh}hk odjhk ickjdlh}
U][email protected]]HU ^PG]CDK
^ckdkjlh}hk }cpigu}h`dod}hu
^ckdkjlh}hk hl}dqd}hu ~hmh ~B ho}cpkh}da
^ckdkjlh}hk hl}dad}hu mhohi lclsh}hk dgk thkj `cp`cmh
^ckdkjlh}hk ~pg}cdk od~h}
^ckspskhk lcpck}hkhk }cpbhmh~ ~pg}cgodudu
Ahpihlgldkc}dl
CLU^PCUD ^PG]CDK
^ckdkjlh}hk }dkjlh} clu~pcud md bgu} kgkkh}spho
]hpjc} clu~pcud lc oglhud ucosohp thkj `cp`cmh
^ckhi`hbhk }hj sk}sl ickmc}clud clu~pcud ~pg}cdk
^cps`hbhk igmdadlhud ~gu}}phkuoh}dgkho
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =
-
7/23/2019 Protein Engineering _ Siti Julaiha
5/98
UDAH])UDAH] OHDK
Ickhi`hblhk }hj sk}sl iciahudod}hud ~cispkdhk
Ickjs`hb lceckmcpskjhk sk}sl ~godicpduhud
Ickhi`hblhkk }hj sk}sl iciqdushoduhudlhk oglhoduhud
Icpclhthuh ickjdlh} ud}su hogu}cpdl
Ickjs`hb }d}dl dugcocl}pdl
Ickspsklhk diskgjckdud}hu
H ODLHUD PCLHTHUH ^PG]CDK
) ^ci`sh}hk ckvdi
) Ici~pgmslud fckdu g`h} g`h}hk
) Ickjbhudolhk qhludk hk}d`gmt }cp}ck}s
@c`cph~h Egk}gb H~odlhud ohkjuskj ucehph lgicpudho uc`hjhd `cpdls} 6
) Ickjjslhkhk ~cphkehb'uehaagom ~pg}cdk `cpmhuhplhk mgihdk ~hmh ^pg}cdk H sk}slickjci`hkjlhk igoclso igocls uc~cp}d hk}d`gmt "Egi~hkt 6 Haad`gmt$
) Ickhi`hblhk huhi hidkg thkj }dmhl }cplgmclhk lh~hmh ~pg}cdk/ iciskjldklhk udk}cuh
~pg}cdk mckjhk mdqcpud}t `hbhk ldidh "Egi~hkt 6 Hi`pr$
) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk ~cp~dkmhbhk sk}sl ickjci`hkjlhk igoclso
igoclso uc~cp}d hk}d`gmt/ ici`sh} ~ckjjh`skjhk ~pg}cdk ~pg}cdk "Asud$ mhk pcec~}gp
hjgkdu} "Egi~hkt6 @dgPcrdu$
) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk oce}dk ]d~c E sk}sl ickjci`hkjlhk igoclso
igoclso uc~cp}d hk}d`gmt/ }clkgogjd ~pg}cdk }pdicpduhud "Egi~hkt 6 @gpchk$+
) Icpclhthuh ~pg}chucu sk}sl ickspsklhk igoclso igoclso thkj md}hpjc}lhk "Egi~hkt6
Eh}hotu}$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 3
-
7/23/2019 Protein Engineering _ Siti Julaiha
6/98
) Ickjjskhlhk mgihdk Ad`pgkcl}dk sk}sl ickjci`hkjlhk igoclso igoclso uc~cp}d hk}d`gmt/
icked~}hlhk ~pg}cdk ~pg}cdk msh askjud mckjhk mgihdk dlh}hk }hpjc} thkj }cpbs`skj
}cpbhmh~ mgihdk ckvdih}dl "Egi~hkt6 Egi~gskm ]bcp~hcs}deu$+
) ^ckjci`hkjhk ~ckdkjlh}hk ic}gmc sk}sl bsihkduhud hk}d`gmt "Egi~hkt6 Lhog`dgu$
) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk od~gehodk sk}sl ickjci`hkjlhk igoclso
igoclso uc~cp}d hk}d`gmt "Egi~hkt6 dcpdu$
) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk Lpdu}ho jhiih sk}sl ickjci`hkjlhk
igoclso igoclso uc~cp}d hk}d`gmt "Egi~hkt6 Uedo$
) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk Udu}cdk Lkg}u sk}sl ickjci`hkjlhk igoclso
igoclso uc~cp}d hk}d`gmt "Egi~hkt6 Ucocegpc$
) Icpclhthuh askjud cacl}gp thkj mddkjdklhk }cpbhmh~ ~pg}cdk ~pg}cdk UID^ uc~cp}d hk}d`gmt
"Egi~hkt 6 ]ps`dgk$+
) Ickjjskhlhk }clkgogjd ^MH sk}sl Icpclhthuh askjud cacl}gp thkj mddkjdklhk }cpbhmh~
hk}d gmt mhk icked~}hlhk ~pg}cdk ~pg}cdk mgihdk kcjh}dqc mhk ~pg}cdk ~pg}cdk mckjhk
~ckdkjlh}hk udah} ~pg~cp}dkth "Egi~hkt 6 Rckegp$+
) Phkehkjhk mhk lgku}pslud mhpd ~pg}cdk h}hs ckvdi `hps mckjhk }clkgogjd `hps'kgqco h}hs
askjud askjud thkj mddkjdklhk mckjhk icigmdadlhud ususkhk huhi hidkg ickjjskhlhk
}clkgogjd pclgi`dkhk mcgluupdkslochdlhudm "PM]$
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 1
-
7/23/2019 Protein Engineering _ Siti Julaiha
7/98
]CLKDL PCLHTHUH ^PG]CDK
Hmh }djh fckdu }clkdl s}hih thkj ucpdkj mdjskhlhk sk}sl icohlslhk pclhthuh ~pg}cdk
~pg}glgo+ Ic}gmc dkd icohlslhk ~ckmclh}hk }cgpd }cpbhmh~ 6
8+ ^ckmclh}hk Mcuhdk Mc Kgqg
0+ ^ckmclh}hk Mcuhdk Phudgkho/ }cpihusl mdhk}hphkth hmhohb }clkdl ~ckmclh}hk
is}hjckcudu Ud}c Mdpce}cm+
5+ ^ckmclh}hk Mcuhdk Is}hjckcudu Hehl'Phkmgi / }cpihusl mdhk}hpkth hmhohb Cqgosud
thkj mdhphblhk'Mdpce}cm Cqgosud/ MKH Ubsaaodkj/ mhk Is}hud Ldidh "]clkdl ^ckmclh}hk
Jodlgudohud/ ^cjdohud/ Od~dmh/ moo$
=+ ]clkdl ^ckjjh`skjhk ~pg}cdk "Asudgk ^pg}cdk$
Sk}sl oc`db fcohukth ucish }clkdl }clkdl dkd hlhk md~h~hplhk uc`hjhd cpdls} 6+
U]PH]CJDU MCUHDK MC KGQG
]clkdl thkj ickhzhplhk lciskjldkhk ~ci`ck}slhk u}psl}sp hps mckjhk ehph 6
) Ickjdmck}dadlhud ususkhk mhk jcgic}pdl huhi hidkg thkj g~}diho sk}sl u}h doduhud
jcgic}pt `hel`gkc
) Ickjci`hkjlhk hojgpd}ih thkj ~gzcpaso sk}sl ugosud g~}diho
) Ickmcuhdk mhk icpclgku}pslud ~pg}cdk udk}c}dl ickjjskhlhk dkapgihud mhpd u}psl}sp 5
mdickud mhpd hlsisohud ~pg}cdk hohi mhk dlh}hk dlh}hk ~pg}cdk+
Egk}gb 6
Ic}bdgkdkc 6 Ici~cpodbh}lhk ucjick pdjdm/ eck}pho ~hp} ucjick bcodr mhk mhcphb `spdcm
kh}spho ~pg}cdk+
]bpcgkdkc 6 Ici~cpodbh}lhk ucjick aocrd`oc
Otudkc 6 Ici`ck}sl ucjick bcodr thkj hdl mhk ickjhfslhkkth ~hmh ]cpidkho)E
Ocsedkc 6 Ici`ck}sl ucjick Bcodecu thkj u}h`do/ ~pcacp `spdcm pcjdgk/ md}cislhk ~hmh
~gudud }ckjhb bcodr
) Ickcislhk ususkhk Huhi hidkg thkj }cp`hdl/ sk}sl icihu}dlhk dlh}hk ~hmh u}psl}sp
thkj md~dodb / uc~cp}d dlh}hk ho~bh bcodr h}hs `c}}h `hppco+
Egk}gb 6 Ickjdmck}dadlhud huhi hidkg thkj }cpmh~h} ~hmh u}psl}sp ~pg}cdk thkj skdl
`cpmhuhplhk ~cps`hbhk ~cpagpihkud ckcpjd }cpigmdkhidl ~hmh phk}hd ~pg}cdk+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc :
-
7/23/2019 Protein Engineering _ Siti Julaiha
8/98
) Mcuhdk lgi~s}hud ~pg}cdk mckjhk hojgpd}ic ickcislhk ususkhk huhi hidkg sk}sl
ickjdmck}dadlhud ususkhk huhi hidkg thkj icidodld ckcpjd ckcpjd pckmhb sk}sl u}psl}sp
}hpjc}+
@hjhdihkh U}psl}sp ~pg}cdk md}ck}slhk
Uc`hjdhk `cuhp u}psl}sp ~pg}cdk thkj mdlckho uhi~hd uhh} dkd }cohb md~cehblhk mckjhk }clkdl
clu~cpdick}ho
R)pht lpdu}hogjphad/
thkj `dhuhkth ici`cpdlhk mh}h pcugosud }dkjjd }c}h~d }dmhl ici`cpdlhk dkagpihud zhl}s
}cpjhk}skj ~hmh aoclud`dod}hu lgkagpihud ~pg}cdk d}s+
KIP "u~cl}pgulg~d pcugkhkud ihjkc}$/
thkj icktcmdhlhk mh}h pcugosud hjhl pckmhb ~hmh sisikth mhk }cp`h}hu sk}sl ~pg}cdk thkj
pcoh}da lcedo/ }c}h~d mh~h} ici`cpdlhk dkagpihud zhl}s thkj `cpjhk}skj }ck}hkj jcphl uc`shb
~pg}cdk mhohi ohps}hk+
R)PHT LPDU]HOGJPHAD
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 2
-
7/23/2019 Protein Engineering _ Siti Julaiha
9/98
Udkhp ihusl "mh}hkj mhpd ldpd h}hu$ icktc`h`lhk uc}dh~ ueh}}cpcp icihkehplhk lci`hod
uc`hjdhk lcedo dk}ckud}hu uc`hjhd jcogi`hkj `goh+ Fdlh ueh}}cpcpu mdususk udic}pdu mckjhk
~ciduhbhk m/ jcogi`hkj dkd `goh hlhk ucohphu "}hi`hblhk lgku}psl}da$ bhkth mhohi hphb ihkh
fhohk)~hkfhkj ~cp`cmhhk icpclh 0m mguh uhih mckjhk ush}s iso}d~oc dk}cjcp mhpd ~hkfhkj
jcogi`hkj + Mhohi bho dkd/ `hjdhk mhpd `cplhu thkj ihusl md`cogllhk gocb 0 usms}/
ici~pgmslud }ci~h} pcaoclud
Mdhjphi ^d}h gaitgjog`dk u}psl}sp/ ickskfsllhk bcodlu hoah `cpzhpkh+ ~pg}cdk }cpuc`s} ~hkfhkj/ igoclso
odkdcp mckjhk pd`shk h}gi/ khisk ~gudud pcoh}da uc}dh~ h}gi }cohb md}ck}slhk mckjhk pcugosud us`)h}gi
mckjhk lpdu}hogjphad udkhp)R+ Lhpckh usod} sk}sl iciqdushoduhudlhk ucish h}gi uclhodjsu/ ~d}h
ickskfsllhk fhohk lhuhp mhpd ~godicp ~pg}cdk mhpd K)}cpidksu "`dps$ lc E)}cpidksu kth "icphb$+
U}psl}sp lpdu}ho ~pg}cdk ~cp}hih mhpd idgjog`dk ~hsu u~cpih/ uc`hjhdihkh md}ck}slhk gocb Ihr
^cps}v mhk Lckmpcz Udp Fgbk Egzmcpt ~hmh }hbsk 8432/ thkj icktc`h`lhk Bhmdhb Kg`co
mhohi Ldidh+
Mdaphlud udkhp)R hkhodudu idgjog`dk hzhokth mdig}dqhud gocb g`ucpqhud lpdu}ho idgjog`dk md lgohi
lcpdkj mhpd mhphb md h}hu jcohmhl lh~ho mhpd ~cp`spshk ~hsu+
KIP "U^CL]PGLG^D PCUGKHKUD IHJKC]$
KIP hmhohb ~ckjc}hbshk u}sl}sp `dgogjd/ thkj mdh~hodlhudlhk gocb KIP sk}sl ickjdkqcu}djhud
fckdu ~pg}cdk ~pg}cdk+
^pg}cdk KIP mdohlslhk ~hmh uhi~co hdp mhpd ~pg}cdk thkj uhkjh} ispkd+
Uhi~co }cpmdpd mhpd hk}hph 5>> mhk 1>> idepgod}pcu mckjhk lgkuck}phud ~pg}cdk mhohi pck}hkj
>/8)5 idodigo+
Usi`cp ~pg}cdk mh~h} `cps~h hohi h}hs mdbhudolhk mhohi udu}ci clu~pcud ickjjskhlhk }clkdl
MKH pclgi`dkhk icohosd pclhthuh jckc}dl+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 4
-
7/23/2019 Protein Engineering _ Siti Julaiha
10/98
^hkhb `dps iczhldod gpdck}hud K ) B dlh}hk ~c~}dmh g`odjhud thkj md~dodb+ Mckjhk ickck}slhk
gpdck}hud thkj esls~ g odjhud pcoh}da }cpbhmh~ icmhk ihjkc} oshp/ u}psl}sp ~pg}cdk mh~h}
md}ck}slhk
Kslodp ~ckck}shk u}psl}sp ihjkc}dl pcugkhkud ickjbhudolhk uc`shb ckuci`oc u}psl}sp+ U}psl}sp
bhkth hlhk `cp}cis fdlh mh}h thkj esls~ sk}sl ickmdl}c od~h}hk }cp}ck}s+ Mhohi u}psl}sp dkd/
bhkth lhusu sk}sl ickfhmd `hjdhk mhpd u}psl}sp+ Mhpd ^M@ 8UUS+
LC][email protected]]HUHK
U}ph}cjd mcuhdk mc Kgqg dkd icidodld `c`cph~h odid}hud h}hs ~pg`oci uc~cp}d 6
) Lc}cpucmdhhk mh}h u}psl}sp ~pg}cdk 5 Mdickud thkj }cp`h}hu+
) ^ckjc}hbshk thkj }cp`h}hu }cpbhmh~ dois dlh}hk ~pg}cdk/ u}psl}sp }cpbhmh~ askjud `dgogjud
~pg}cdk ~pg}cdk }cpuc`s}+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 8>
-
7/23/2019 Protein Engineering _ Siti Julaiha
11/98
H^ODLHUD U]PH]CJD
Ususkhk skdl ~pg}cdk ickjbhudolhk phkehkjhk }cpbhmh~ ususkhk ^g}ckudho ^pg}cdk mhpd
`c`cph~ ph}su pd`s huhi hidkg mh~h} mdbhudohk {8>85> ususkhk ~pg}cdk ~g}ckudho icohosd
ic}gmc lgi~s}hud h}hs clu~cpdick}ho+6
Cu}dihud ckcpjd ~hmh mcuhdk ~pg}cdk thkj ici~cpodbh}lhk ~cps`hbhk ckcpjd ucohih
~pgucu agomdkj+
Bd~g}cuh }cpigmdkhidl thkj mdisohd mhpd u}psl}sp huod ~pg}cdk thkj md`cpdlhk mckjhk
hmhkth lgkagpihud huhi hidkg thkj icidkdiholhk ckcpjd `c`hu igoclso+ Bhudo mhpd
cu}dihud ckcpjd dkd ici~cpodbh}lhk 6
g Bg} u~g} pcudms sk}sl hmhkth dk}cphlud ckcpjd
g Mh}h U}h}du}dl }cpbhmh~ askjud cacl}da ckcpjd "UCCAu$ thkj `cpihkahh} sk}sl mh}h`huc
u}psl}sp ~pg}cdk
g Udah} udah} adudl }cpbhmh~ askjud cacl}da ckcpjd "^CCAu$ thkj `cpihkahh} sk}sl
icohlslhk hkhoduh lgi~s}hud+
g Udah} udah} ci~dpdu }cpbhmh~ askjud cacl}da ckcpjd "CCCA$ thkj `cpihkahh} sk}sl
ickjhkhoduh udkjoc }cpigmdkhidl mhk iso}d~oc ud}c is}hud+
g Lcu}h`dohk ~pg}cdk thkj `cpjskh sk}sl icktsusk `hpdu iso}d~oc dlh}hk mhk mhcphb
lgkucpqh}du ~hmh fckdu ~pg}cdk "Esppck} G~dk / dk U}pse}spho @dg+ 0>>0/ 806==8$
Mcuhdk ~pgucu lgi~s}hudgkho fsjh `cpjskh sk}sl icphkehkj igmco agomdkj dkqcpuc
`hjdhk ~pg}cdk lcmhohi phkehkjhk sk}hd thkj mdhusiudlhk uc`hjhd pg}hicp iso}d~oc mhk
~cphkehb }c}h~+ Pg}hicp thkj `cp`cmh mhpd }pd}g~hk huhi hidkg ~hmh ~gudud }cp}ck}s
mhohi ~pg}cdk+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 88
-
7/23/2019 Protein Engineering _ Siti Julaiha
12/98
Igmco mcuhdk agomdkj dkqcpuc "H$ ~hp}ud ~pg}cdk mdmcudjk udmc)ebhdku ickjhusihudlhk iso}d~oc pg}hicp "icphb$mhk uehaagom }c}h~ "`dps$+ "@)M$ pg}hicp `cp`cmh }pd}g~hk huhi hidkg ~hmh ~gudud }cp}ck}s md ~pg}cdk+
Ickjdmck}dadlhud ususkhk mhk jcgic}pdl huhi hidkg thkj g~}diho mckjhk }hpjc}
}cpbhmh~ u}h`doduhud jcig}cpd `hel`gkc ~pg}cdk+
g
Mcuhdk lgi~s}cp sk}sl ahl}gp }sigp kclpgudu "]KA$ qhpdhk ~hmh mc~de}cm jgom+ Is}hud/ icphb/ ickecjhbmcuhdk qhpdhk mhpd dlh}hk ]KA pcuc~}gp/ `dps+ ^ckhi`hbhk is}hk bhudo ]KA ~hmh ehi~sphk bc}pgjck}pdicp/ icidodld hl}dqd}hu thkj mdlgi~pgiducm+ ^ckhi`hbhk is}h} `cpoc db ]KA ickfhkfdklhk ocqco pckmhbgig}pdicp hl}dqc kh}dqc ]KA+ Jhi`hp `cpmhuhplhk ~hmh ih}cpdho us~ocick pcacpckec 54+ U}psl}sp lpdu}ho8]KP mdjskhlhk sk}sl icked~}lhk pc~pcuck}h}da u}psl}sp+
U]PH]CJDU MCUHDK PHUDGKHO ^PG]CDK
Icps~hlhk }clkdl thkj ickjjskhlhk ~ckjc}hbshk uedck}dadl mhk ickmc}hdo }ck}hkj oh}hp
`cohlhkj uc~cp}d u}psl}sp mhk askjud ~pg}cdk h}hs udah} ~pg}cdk/ ucp}h ususkhk jck thkj hlhk
mdpclhthuh ~hmh ususkhk ~god~c~}dmh sk}sl mdjskhlhk mhohi6
) Ickmh~h}lhk ~pgmsl ~pgmsl jck thkj `hps mhpd ~pg}cdk thkj hmh
) Ickcislhk bs`skjhk hk}hph u}psl}sp mhk askjud ~pg}cdk ucp}h u}h`dod}hu Igoclso icohosddkagpihud mhpd u}psl}sp 5 Mdickud ~pg}cdk+
) Ickjdmck}dadlhud ususkhk Huhi Hidkg uc~cp}d ickjc}hbsd huhi hidkg thkj icidodld
ckcpjd }cpckmhb sk}sl icps`hb u}psl}sp dlh}hkkth+
) Ickjci`hkjlhk ~ckjc}hbshk ~ckjc}hbshk `hps }ck}hkj lhphl}cpdu}dl ~pg}cdk+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 80
-
7/23/2019 Protein Engineering _ Siti Julaiha
13/98
) Icphkehkj ~pg}cdk h}hs ckvti mckjhk udah} thkj mddkjdklhk icohosd }clkdl ihkd~sohud
`cpmhuhplhk ~ckcph~hk h}hs ~ckdpshk }clkdl is}hud hohi uc~cp}d 6
) Ud}c Mdpce}cm Is}hjckcudu mckjhk ^EP)@hucm h}hs Cppgp pgkc
) Is}hud ^ckhi`hbhk h}hs ^ckjbdohkjhk mckjhk iso}d~odlhud ^EP `hucm
ckvdi kseochuc mhk ' h}hs Logkdkj+
) ^cps`hbhk Mgihdk h}hs ^ckjjh`skjhk mgihdk "Asudgk$ mckjhk Ckvdi
Odjhud/ mhk iso}d~odlhud EP)@hucm/ moo
Ic}gmc dkd ismhb mddi~ocihk}hudlhk mhk ucehph sisi }dmhl ici`s}sblhk `dhth `cuhp/ ucfhl
}clkdl ~ckmclh}hk ud}c mdpce}cm is}hjckcudu md}cph~lhk mckjhk `hdl+
LC][email protected]]HUHK
U}ph}cjd Phudgkho mcuhdk fsjh icidodld `c`cph~h odid}hud h}hs ~pg`oci uc~cp}d 6
) ^ckjc}hbshk }ck}hkj dois dlh}hk ~pg}cdk/ fsjh }cpbhmh~ askjud `dgogjud/ h}hs u}psl}sp ~pg}cdk
uhkjh} }cp`h}hu mhk ucpdkj }dmhl }cpucmdh
) Fdlh mdh~odlhudlhk/ihlh ici~cpldphlhk cacl cacl mhpd qhpdhud is}hudkth hlhk uhkjh} usod}
mdohlslhk+
) U}ph}cjd mcuhdk Ud}c Mdpcl}cm Is}hjhkcudud icidodld odid}hud h}hs ~pg`oci mckjhk
@hkthlkth od`phpd ~hmh is}hk thkj mdbhudolhk
H^ODLHUD MCUHDK PHUDGKHO
H~odlhud ]clkdl }clkdl dkd m~cposlhk mhohi `cp`hjhd bho uc~cp}d 6
) Icohlslhk hkhoduh }cpbhmh~ dk}cphlud hk}hph ~pg}cdk ~pg}cdk/ mhk agomdkj'skagogmdkj ~pg}cdk
) Icohlslhk ~cpigmcohk u}psl}sp ~pg}cdk+
) Icohlslhk hkhoduh }cpbhmh~ dk}cphlud dlh}hk MKH/
) Icohlslhk clu~ogphud ususkhk ~pg}cdk/
) Icohlslhk lgk}psuld bd`pdm ~pg}cdk/ ~pg`dkj ~pgig}cp mhk
) Ickjclu~pcudlhk pg}cdk mhk Pcjsohud pcjdgk ucp}h
) Ickjhkhoduh hl}dad}hu ckvdi+
) Icohlslhk igmdadlhud udah} udah} ldidh ~pg}cdk
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 85
-
7/23/2019 Protein Engineering _ Siti Julaiha
14/98
) Ici~pgmslud g`h} mckjhk iduhokth ickjcacl}dalhk Pd`gkslcohud thkj mdjskhlhk ~hmh
}cph~d }sigp+
) ]clkdl dkd mh~h} ickjbhudolhk h~odlhud mhohi ickjlhphl}cpdu}dl fckdu hk}d`gmd+
) ^ckcod}dhk thkj clu}ckuda mhk u}smd }ck}hkj mhcphb dl}hk >0 "odjhkm$ ~hmh ~pg}cdk
bcigjog dk+ Mckjhk u}psl}sp bcigjog`ck }cpohps} ~hmh lpdu}hogjphad R)pht / huhi hidkg
thkj `cp}sjhu sk}sl ickjdlh} odjhkm ~hmh ~pg}cdk/ `cpuhih mckjhk bcodr u~cudadl mhk
~gududkth md}ck}slhk mhk mh~h} md}hi~dolhk+ Pcudms pcudms thkj `clcpfh ~hmh lgkfsklud
mckjhk edkedk bcic ~gp~dpdk+
Jhi`hp ici~cpodbh}hlk }c}phicp bcigjog`dk mckjhk ci~h} mhcphb thkj ickjdlh} odjhk mckjhk mdu}ho mhk~pgludiho bdu}dmdkcu mhohi igmco }gkjlh} `dps ecphb+ Edkedk bcic ~gpadpdk md}hi~dolhk mhohi igmco }gkjlh}icphb mhk ~s}db+
Jhi`hp dkd ici~cpodbh}lhk ~cocmhlhk md lhzhuhk thkj ickjdlh} odjhk+ Phk}hd uhi~dkj mhpd huhi hidkg huhihidkg lsked ~hmh dlh}hk odjhk "G0 mhohi jhi`hp$ mhk edkedk bcic ~gp~dpdk md}skfsllhk mhohi igmco }gkjlh}+
Pshkj lskdkj ~cksb igoclso icps~hlhk bcodlu ucld}hpkth thkj ici`ck}sl bcigjog`dk+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 8=
-
7/23/2019 Protein Engineering _ Siti Julaiha
15/98
Ickjjskhlhk u}ph}cjd mcuhdk phudgkho mh~h} ickthphklhk is}hud thkj iskjldk sk}sl
huhi hidkg lsked thkj `cp}hkjjskj fhzh` h}hu ~ckjdlh}hk bcigjog dk ~hmh
odjhk+ Uc`hjhd egk}gb/ fdlh mddkjdklhk u}cpdehoot ickjbhohkjd ud}su ~ckjdlh} fdlh bcigjog`dk/
mdpckehkhlhk ici`sh} ud}su)is}hud sk}sl ici~cplckholhk is}hud thkj hlhk ici`cpdlhk
udah} adudl mhk cocl}pgldidh thkj mdlcbckmhld+ Icidodb ackdohohkdk sk}sl ickjjhk}dlhk ocsudk
~hmh ~gudud 04 hlhk ickjdud ud}su mckjhk phk}hd uhi~dkj hpgih}de thkj `cuhp+ Bhudo dkd
mh~h} mdodbh} ~hmh jhi`hp `cpdls} H mhk @+
Jhi`hp H+ ickjjhi`hplhk mhcphb thkj ickjdlh} h}hsuhls mhpd us`skd} bcigjog dk+ U~hud zhpkh bdfhs mddudmgihdk ici~cpodbh}lhk pcudms ocsudk zdom }t~c mhkgpdck}hudkth ~hmh ~gelc}+ Igoclso `cpzhpkh icphb hmhohbdludjck mhk md}hi~dolhk `cpdlh}hk }cpbhmh~ jps~ bcic
~gpadpdk "igoclso aoh} ~hmh igmco }gkjlh}$+
Jhi`hp @+ ici~cpodbh}lhk mhcphb thkj ickjdlh}uc}cohb ocsudk zdom }t~c }cohb mdjhk}dlhk gocb uc`shbackdohohkdk+ U}ph}cjd phudgkho ackdohohkdk ickjdud uhlsbcigjog`dk mdlgkadpihud ~hmh jhi`hp thkj mdmhuhplhkmhpd r)pht lpdu}hogjphad+
U]PH]CJDU MCUHDK PHUDGKHO U]PSL]SP MHK ASKJUD ^PG]CDK
U}smd Igmcpk pcugosud thkj oc`db }dkjjd Dkd hmhohb ic}gmc clu~cpdick}ho sk}sl mh~h} icides
od~h}hk uhi~co skagomcm ~pg}cdk/ mhk ickjhih}d mdkhidlh thkj mdbhudolhk+
]clkdl ec~h} mdjskhlhk ucehph oshu }cpihusl kcs}pgk bhi`sphk/ so}phahu}/ ~ckehi~sphk
ugosud/ ag}gldidh ic}gmc/ mhk u~cl}pgulg~d ohucp usbs icogi~h}+
Lgi~s}hud fsjh ici~pcmdlud u}psl}sp }cpudcp ~pg}cdk+
]clkdl dkd}dg Mc kgqg "ici~pcmdlud u}psl}sp ~pg}cdk `cpbs`skjhk mckjhk lgi~s}hud$/ Uhkjh}
`cp`cmh mckjhk Ic}gmc od~h}hk Mdkhidlh Igoclso ~pg}cdk+
"IM$ icps~hlhk ~cphkjlh} ~ck}dkj ici~cohfhpd ~cod~h}hk ~pg}cdk mhk mdkhidlh md udodeg+
Lhpckh `dhth lgi~s}hud/ udisohud IM od~h} mckjhk hdp clu~odud} bhkth }cp`h}hu ~hmh ~c~}dmh
mhk ~pg}cdk thkj uhkjh} lcedo+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 83
-
7/23/2019 Protein Engineering _ Siti Julaiha
16/98
IM udisohud ~pg}cdk thkj oc`db `cuhp }c}h~ }cp`h}hu ~hmh mdkhidlh u}psl}sp clu~cpdick}ho h}hs
`cpohkjuskj ~hmh }ci~cph}sp }dkjjd+
Sk}sl ickudisohudlhk ~pguccu od~h} thkj ohih "mdoshp ucld}hp 8 idlpgmc}dl$/ uc~cp}d ~pg}cdk
od~h} slsphk lcedo "ucld}hp 3> pcudms$ h}hs oc`db `cuhp/ `c`cph~h ~ckmclh}hk h}hs
~cktcmcpbhkhhk mhohi igmco ~pg}cdk ~cpos md~cplckholhk+
]clkdl sk}sl ici~cohfhpd pg}cdk Agomdkj
Edpesohp Mdebpgdui U~cl}pgulg~d
hmhohb uhohb uh}s hoh} ~hodkj sisi mhk mhuhp sk}sl ickjslsp ~cktcph~hk ehbhth
}cp~gohpduhud udplsohp+
Mhohi ~pg}cdk/ u}psl}sp uc~cp}d oci`hphk bcodedcu ho~bh mhk `c}h thkj ldpho/ mhk mckjhk
mcidldhk icktcph~ ehbhth }cpuc`s}+
^cktcph~hk ehbhth dkd `cp}dkmhl uc`hjhd ~ckhkmh }dkjlh} agomcmkcuu mhpd hkuhi`co ~pg}cdk+
]clkdl mdjskhlhk sk}sl ickjslsp lcucdi`hkjhk ~pg}cdk mckjhk ickjslsp ~cps`hbhk
ucph~hk uc`hjhd askjud lgkuck}phud mckh}sphk} h}hs usbs+
Fckdu u~cl}pgulg~d fsjh mh~h} mdlgi`dkhudlhk mckjhk ~cphkjlh} ec~h})~ckehi~sphk/ sk}sl
ickjslsp ~pg}cdk ldkc}dlh od~h} mhk sk}sl ickjbhudolhk ~og} ebcqpgk+
^cplci`hkjhk mdebpgdui odkjlhphk jc}hphk "QEM$ icod h}lhk Agspdcp ]phkuagpi "AA]$
dku}psick/ sk}sl ickck}slhk lgkagpihud ~pg}cdk mhohi ohps}hk `hblhk sk}sl igoclso ~pg}cdk
uhkjh} `cuhp+
QEM u}smd ~pg}cdk ucpdkj mdlgi`dkhudlhk mckjhk mdaphlud udkhp R)lpdu}ho ~pg}cdk/ A])DP mh}h
sk}sl ugosud ~pg}cdk mhohi hdp `cph} "m0G$/ h}hs ~cpbd}skjhk lshk}si h` dkd}dg sk}sl
ici`cpdlhk }sjhu u}psl}spho hi`djs thkj }hl m~} md~cpgocb mhpd EM
U~cl}pgic}pd Edpesohp Mdebpgdui "EM$
Hoh} dkd md~cposlhk mhohi icohlslhk ~ckjslsphk u}psl}sp uclskmcp ~pg}cdk mckjhk
ickjjskhlhk }clkgogjd u}hkmhpm mhohi ickjslsp hl}dad}hu g~}dlho ~pg}cdk/ thd}s ickjslsp
~cp`cmhhk h`ugp`hkmd ~hmh u}psl}sp udplsohp `hjdhk lhkhk mhk ldpd u}psl}sp ~pg}cdk icohosd
^gohupduh ehbhth mhpd uhi~oc ~pg}cdk+
Lc}c}h~hk u}hkmhp Ahp)SQ h}hs mhcphb hidmc hmhohb 8:> )03> ki sk}sl dlh}hk ~c~}dmh mhk
mhcphb Kchp SQ hmhohb 03> 5>> ki sk}sl hpgih}dl huhi hidkg+ Uc~cp}d thkj }cpodbh} ~hmh
jhi`hp md `hzhb dkd+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 81
-
7/23/2019 Protein Engineering _ Siti Julaiha
17/98
Clu~cpdick }clkdl ~ckck}shk u}psl}sp ~pg}cdk
Mdod~h} u}psl}sp ~pg}cdk ucehph ps}dk md}ck}slhk gocb Lpdu}hogjphad udkhp)R mhk KIP+
Icihbhid mdebpgdui icodkjlhp6 mhuhp)mhuhp ~gohpduhud sk}sl `ckhp)`ckhp icihbhid
mdebpgdui odkjlhphk/ thkj ~cp}hih bhpsu icihbhid mhuhp)mhuhp ~gohpduhud
]cp~gohpduhud odkdcp ehbhth hmhohb ehbhth thkj gudohud }cp`h}hu sk}sl ~cuhzh} }skjjho+ Ucish
kcjhph }cp~gohpduhud ehbhth mh~h} mdjhi`hplhk uc`hjhd fsiohb mhpd odkchp }cp~gohpduhud }cjhl
ospsu uh}s uhih ohdk/ `dhuhkth mdpsfsl lc ~ckhi~do uc`hjhd qcp}dlho mhk bgpdvgk}ho }cp~gohpduhud
ehbhth+ Dkd md}hi~dolhk mhohi hkdihud md `hzhb dkd+ "Jcphllhk igsuc md h}hu jhi`hp sk}sl
icodbh} hkdihud Hkdihud md`sh} mhpd ~pgjphi Cihkdi++$
Lhohs iduhokth ld}h hi do bgpdugk}ho mhk qcp}dlho }cp~gohpduhud hi~od}smg jcogi`hkj ehbhth
thkj uhih md ahuc uh}s uhih ohdk/ jcogi`hkj ehbhth thkj mdbhudolhk "`dps$ hmhohb odkchp
}cp~gohpduhud ~hmh =3 mcphfh}/ uc~cp}d }cpodbh} ~hmh hkdihud md hzhb+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 8:
-
7/23/2019 Protein Engineering _ Siti Julaiha
18/98
Fdlh lcmsh ~gohpduhud thkj lcoshp mhpd ahuc/ jcogi`hkj pcuso}hk `cpbck}d ickfhmd }cp~gohpduhud
odkdcp+ Iduhokth/ fdlh uhohb uh}s }cp~gohpduhud hmhohb lcoshp mhpd ahuc mckjhk thkj ohdk gocb
jcogi`hkj)lshp}ho/ pcuso}hk hlhk bcodlu mhk mdlckho uc`hjhd ehbhth }cp~gohpduhud udplsohp
"E^O$+ ]bc bcodlu mh~h} `cps~h)}hkjhk lhkhk "P)E^O$ h}hs ldmho "O)E^O$ mhk kgk)jhi`hp
ecpidk us~cpdi~guh oc+
Skusp g~}dl thkj ickjlgkqcpud hk}hph ehbhth }cp~gohpduhud odkdcp mhk ehbhth }cp~gohpduhud
udplsohp mduc`s} ~dpdkj uc~cpci~h})jcogi`hkj+ Uc`shb ~dpdkj uc~cpci~h})jcogi`hkj
`dpcapdkjck}/ thd}s dkmclu `dhu mdodbh} gocb bgpdvgk}ho mhk qcp}dlho }cp~gohpduhud ehbhth `cp`cmh+
Uc`shb ~coh} ucushd `cpgpdck}hud hlhk ickjlgkqcpud ehbhth }cp~gohpduhud odkdcp lc udplsohp
ehbhth }cp~gohpduhud mckjhk ici~cpohi`h} uhohb uh}s lgi~gkck odkdcp `hogl ucbs`skjhkmckjhk thkj ohdk ucbdkjjh icpclh lcoshp uc~cpci~h})jcogi`hkj ahuc+ Dkd hlhk ickjbhudolhk
uc`shb `cplhu `hdl)ldpd h}hs lhkhk E^O+
Lhohs iduhokth ld}h hi do bgpdugk}ho mhk qcp}dlho }cp~gohpduhud hi~od}smg jcogi`hkj ehbhth
thkj uhih md ahuc uh}s uhih ohdk/ jcogi`hkj ehbhth thkj mdbhudolhk "`dps$ hmhohb odkchp
}cp~gohpduhud ~hmh =3 mcphfh}/ uc~cp}d }cpodbh} ~hmh hkdihud md hzhb+
U~cl}pgic}pd Aosgpguckuc
Thd}s hoh} thkj mdjskhlhk sk}sl icohlslhk ~ckjslsphk thkj uci~spkh mhohi
ickjlgkadpihud ~cps`hbhk ~cps`hbhk thkj }cpfhmd ~hmh ~pg}cdk+
Uc~cp}d thkj mdohlslhk ~hmh ~ckjoh`cohk Ucpg}gkdk K hec}do}phkuacphud "HHKH]$ mckjhk
aosgpcuck "AO$ mhk pbgmhidk "Pb$ / thkj ickjbhph~lhk }cpfhmdkth }phkuacp pcugkhkud ckcpjd
aosgpcuck "APC]$ sk}sl HHKH] uc`hjhd igkgicp "kg APC]$ mhk uc`hjhd godjgicp "APC]$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 82
-
7/23/2019 Protein Engineering _ Siti Julaiha
19/98
H}hs fsjh ~coh`cohk dk qdqg ~hmh Jos}h}dgkc U)}phkuacphud "JU]$ thkj ickjhkmskj pcudmsc
udu}cdk K)}cpidkho mckjhk ici`phk ~cpidh oc }c}phic}dopbgmhidk }bdgcu}cp "]IP$+
]CLKDL MCUHDK PHUDGKHO ^PG]CDK MCKJHK ^CKMCLH]HK
IS]HJCKCUDU UD]C MDPCE]CM
]clkdl Is}hud Ud}c Mdpcl}cm ickdps is}hud hohi thkj }cpfhmd ~hmh jck gpjhkduic bdms~ mckjhk
icps`hb pcudms huhi hidkg }skjjho icohosd }clkdl is}hud lgmgk thkj ickjlgmclhk huhi
hidkg+
]clkdl dkd hmhohb ic}gmc }cpohih thkj mdjskhlhk ~cp}hih lhod mhohi pclhthuh ~pg}cdk+
Mckjhk udk}cuh godjgkslocg}dmh thkj mdihusslhk lc mhohi ush}s ~ohuidm+
Ic}gmc ~cp}hih mhpd ud}c mdpce}cm is}hjckcdu mdlci`hkjlhk mcjkh ic}gmc Udkjoc ^pdicp/ gocb
Jdoodhi c}+ho 842> mhk Vhoocp mhk Uid}b/ 8425+ Uc~cp}d thkj mdmculpd~udlhk ~hmh ic}gmc hohi
thkj icod`h}lhk udk}cuh dk qd}pg MKH mckjhk udk}cuh ldidh Godjgkslocg}dmh ": bdkjjh 0> phk}hd
~hkfhkj kslocg}dmh$ thkj ici`hzh `huc thkj ucushd mckjhk ususukhk thkj mdockjlh~dkth+
Ic}gmc dkd ici`s}sblhk MKH thkj }cpucmdh ~hmh `ck}sl phk}hd }skjjho sk}sl mdis}hudlhk/
mhk mdlogkdkj mckjhk qcl}gp jckc I85 sk}sl ici~cpismhbkth+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 84
-
7/23/2019 Protein Engineering _ Siti Julaiha
20/98
^cp`hdlhk ^pg}cdk mckjhk ic}gmc MKH Iduih}eb dkd hmhohb uhohb uh}s ~pgucu thkj ~hodkj
~ck}dkj sk}sl ickjlgpclud lcuhohbhk pc~odlud mhohi gpjhkduic bdms~/ ucbdkjjh mh~h}
ickdkjlh}lhk lchlsph}hk pc~odlhud MKH bdkjjh ickeh~hd ahl}gp 8>>>+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 0>
Ususkhk Huod Huhi Hidkg
Udock} Is}hud h}hs Iduuckuc thd}s
~cps`hbhk ususkhk kslocg}dmh }hk~h
hmhkth ~cps`hbhk ususkhk ~pg}cdk+
Kgkuckuc thd}s ^cps`hbhk ususkhk
huhi hidkg thkj icps`hb }cpidkho
h}hs ickjbck}dlhk sfskj)E ~pg}cdk+
^ckhi`hbhk ususkhk huhi hidkg
thkj icps`hb }cpidkho h}hs sfskj)E
~pg}cdk+
^ckjsphkjhk ususkhk huhi hidkg
thkj icps`hb }cpidkho h}hs sfskj)K
~hmh ~pg}cdk
-
7/23/2019 Protein Engineering _ Siti Julaiha
21/98
Udk}cuh ~pdicp MKH `cpudk}cuh mckjhk godjgkslocg}dmh mdihusllhk lc mhohi igoclso
bc}cpgms~ocr mdbhudolhk+ Uc}cohb }phkuagpihud mhpd bgu} C+ egod/ bc}cpgms~ocr dkd ickdi`solhk
bgigms~ocrcu thkj sps}hkkth `cphuho mhpd zdom)}d~c MKH h}hs thkj ickjhkmskj mhuhp
`cpis}hud+ Apclsckud is}hud logk thkj iskeso oc`db pckmhb md`hkmdkjlhk mckjhk logk zdom
}t~c+
^ckjhi`dohk is}hk/ mdohlslhk mckjhk ~ckthpdkjhk bd`pdmduhud huhi kslochm mhk ~pg`c kth
thkj mdoh`co mckjhk 50^)oh`co godjgkslocg}dmh+
^hphic}cp thkj `cp~cphk }cpihusl mdhk}hphkth usbs mhk lgkuck}phud lh}dgk thkj mdmh~h}
uc`hjhd udktho ~gud}da mhpd Is}hud Logk+ Bho dkd iciskjldklhk lcudh~hk sk}sl ickmc}clud
is}hk thkj mddkjdkhlkh "Zhoohec c}+ 8428/ ]ph`gkd c} ho+ 8425$+
Uhkjh} mdhkfsplhk sk}sl icicpdluh sps}hk is}hk ucehph ohkjuskj mckjhk ickjjskhlhk
ic}gmc uclsckudkj MKH/ sk}sl icihu}dlhk `hbzh ~pgucmsp }dmhl icohlslhk ~cps`hbhk
hmqck}da ohdkkth+ Bh Dkd uhkjh} md`s}slhk ~hmh qcpud ~cp}hih }clkdl thkj ickjjskhlhk MKH
~godicphud C)Egod+
^ckjjskhhk thkj MKH ~godicphuc bdjb admcod}ct thkj oc`db `hps mh~h} icidkdiholhk ihuhohb
is}hud hudkj ucp}h ici~cp~ckmcl zhl}s sk}sl ickthodk sk}hd lcmsh+ Ucohdk d}s/ ~godicphuc
dkd }dmhl ickjjhk}dlhk)sk}hd godjgicp/ uc`shb ~pgucu thkj hlhk ickjbdohkjlhk
godjgkslocg}dmh is}hk huod+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 08
-
7/23/2019 Protein Engineering _ Siti Julaiha
22/98
MKH thkj mdlogkc ~hmh ~ohuidm mhhk mdmh~h}lhk mhpd `ck}sl ms~ocr mh~h} fsjh mdlgkqcpudlhk
h}hs mdps`hb ickfhmd igoclso ~hp}dho udkjoc thkj ucushd / mdohlslhk gocb Mho`hmdc)Ie Ahpohkm c}
ho+8420+ Ucbdkjjh mhohi ~ckjci`hkjhkth }cplhmhkj mdjskhlhk fsjh mgs`oc uc`hjhd jhk}d
phk}hd ~pdicp MKH uc~cp}d jhi`hp `hzhb lhkhk+
^CKMCLH]HK ]CLKDL IS]HUD ^[email protected] H]HS
^CKJBDOHKJHK MCKJHK ^EP @HUCM CKVDI KSLOCHUC
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 00
-
7/23/2019 Protein Engineering _ Siti Julaiha
23/98
Uc`shb qhpdhud mhpd ~pgucmsp thkj }cohb mdfcohulhk uc`cosikth mdh}hu/ }clkdl is}hud ~hmh dkd
icod`h}lhk godjgkseocg}dmcu thkj ickjhkmskj ususkhk thkj mh~h} md}hi`hblhk h}hs
mdbh~sulhk+ Ucohih bd`pdmh u}h`do }cp`ck}sl mckjhk zdom }t~c MKH `cpsk}hd }skjjho/ ~pdidkj
udk}cudu MKH dk qd}pg mh~h} }cpfhmd/ hlbdpkth ickdi`solhk logk ucushd mckjhk ususkhk thkj
md}hi`hbhlhk h}hs mdbh~sulhk "Zhoohec c} ho+ 842>/ Kgpphkmcp c} ho+ 8425$+
]CLKDL ^CKMCLH]HK IS]HUD ^[email protected] MGIHDK H]HS
^[email protected] MGIHDK "ASUDGK$ MCKJHK CKVDI ODJHUD MHK
ISOD]D^ODLHUD ^EP)@HUCM
IS]HJCKCUDU THKJ MDHPHBLHK MCKJHK GODJGKSLOCG]DMH MCKJHK
^[email protected] MGIHDK
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 05
-
7/23/2019 Protein Engineering _ Siti Julaiha
24/98
Uc`hjhd egk}gb cpdls} 6
Is}hjckcudu dk Qd}pg
Logkdk jck mh~h} `cpis}hud ucehph lbsusu/ ucbdkjjh iciskjldklhk hmhkth is}hud }cp}ck}s+
Lgi~gkck s}hih thkj md`s}sblhk mhohi ~pg}gego Mdpcl}cm is}hjckcudud dkd mdhk}hpkth 6
) MKH ]ci~oh}c/ ^pdicp Is}hud/ MKH ^godicphuc/ mK]^+ Mhk `c`cph~h qhpdhud ohdk sk}sl
ickdkjlh}lhk cadudckudkth+
H^ODLHUD ]CLKDL ^CKMSLSKJ UD]CM MDPCE]CM IS]HJCKCUDU
PCHLUD PHK]HD ^GODICP ' ^GOTICP EBHDK PCHE]DGK'^EP
]clkdl ^EP icps~hlhk ehph sk}sl ickjdugohud aphjick MKH ucehph ec~h} mhk fsjh sk}sl
ickeh~hd hlsisohud clu~gkckudho mhpd }hpjc} MKH+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 0=
-
7/23/2019 Protein Engineering _ Siti Julaiha
25/98
]clkdl dkd mdmhuhplhk ~hmh sps}hk MKH thkj }cohb md}ck}slhk uc`cosikth/ uc~cp}d
godjgkslog}dmh ~ckmcl "{ 0>`~$ mdlci`hkjlhk mhk mdockjlh~d sk}sl icktsusk MKH }hpjc} thkj
mdh~d} + Godjgkseocg}dmcu `cp}dkmhl uc`hjhd ~pdicpu sk}sl ickthodk MKH thkj ucps~h "Pc~odlhud
MKH$+
Uc}dh~ udlosu pc~odlhud ickjjhkmhlhk fsiohb }hpjc} MKH 6
Lcismdhk hi~odadlhud'ici~cp`hkthl MKH ucehph clu~gkckudho mdohlslhk 6
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 03
-
7/23/2019 Protein Engineering _ Siti Julaiha
26/98
@c`cph~h egk}gb ^EP thkj mh~h} sk}sl Pclhthuh ^pg}cdk mdhk}hphkth 6
) ^EP Icmdh Is}hjckcudu
Is}hjckcudu Godjgkslocg}dmh iduih}eb mh~h} icked~}hlhk }d}dl is}hud thkj mddkjdklhk ~hmh
mhcphb thkj }cohb md}ck}slhk uc`cosikth "ud}c ~pc)mc}cpidkcm$ mckjhk igoclso MKH logkc+
Uhohb uh}s mhpd `cp`hjhd ic}gmc is}hjckcudud hmhohb uco)`huc godjgkslocg}dmh iduih}eb
"sk}sl ho}cpkh}dqc EP)`huc ud}c mdpce}cm is}hjckcudu$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 01
-
7/23/2019 Protein Engineering _ Siti Julaiha
27/98
^ckjjskhhk is}hjckc}dl godjgkslocg}dmc ucehph ohkjuskj ~hmh kslocg}dmh }skjjho thkj
mdus`u}d}sud ~hmh jckc+
Jckc mdlogkdkj ~hmh I85 sk}sl icked~}hlhk pclgi`dkhud MKH phk}hd }skjjho+
^pdicp godjgkslocg}dmh mdphkehkj sk}sl icockjlh~d ucehph `cpsps}hk `hjdhk mhpd ususkhk
jck thkj icod~s}d kslocg}dmh thkj hlhk mdis}hud "H$ mhk ickjhkmskj `huc kgk)
lgi~ocick}hpd thkj mddkjdklhk ~hmh ~gududkth "E$+
Iculd~sk `cps~h dk}cpkho iduih}eb/ hkkchodkj mhpd ~pdicp is}hjckde mh~h} mdohlslhk / mhk
udk}cuh phk}hd lcmsh mh~h} md~cp~hkfhkj mckjhk ~godicphud MKH ucp}h ecohb'jh~ md}s}s~
mckjhk MKH odjhuc+
Bhudo bc}cpgms~ocl mh~h} md}phkuagpihudlhk lc mhohi C)egod/ mdihkh msh ~g~sohud
pclgi`dkh mh~h} mdlci`hodlhk ickfhmd zdomc }t~c mhk is}hk} bgigms~ocl/ thkj lcismdhk
mh~h} mddmck}dadlhudlhk mckjhk bd pdmduhud igocesohp "mckjhk ickjjskhlhk ~pdicp
is}hjckde uc`hjhd ~pg`c godjgkslog}dmh hoco u~cudadl$+
) ^EP @huc Ud}c Mdpce}cm Is}hjckcudu
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 0:
-
7/23/2019 Protein Engineering _ Siti Julaiha
28/98
) ^EP @huc ]spskhk ^pdicp Ud}c Mdpce}cm Is}hjckcudu
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 02
-
7/23/2019 Protein Engineering _ Siti Julaiha
29/98
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 04
-
7/23/2019 Protein Engineering _ Siti Julaiha
30/98
) ^EP Is}hjckcudu
"H$ Is}hjckcudu sfskj 3
^pdicp mdigmdadlhud ~hmh sfskj 3 sk}sl icisohd ~pgucu/ uc`hjhd egk}gb/ Jpgs~ thkj
}cohb mdoh`colhk/ ususkhkkth ickjhkmskj ud}c pcu}pdlud thkj ucushd/ h}hs ~pgig}cp~bhjc sk}sl ickjjcphllhk clu~pcud jck+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 5>
-
7/23/2019 Protein Engineering _ Siti Julaiha
31/98
"@$Is}hjckcudu Ud}c U~cudadl
Is}hjckcudu ickskfsllhk mh~h} ickjbhudolhk ~pgmsl hi~odadlhud mckjhk
ickjoglhudlhk is}hud ~pc)mc}cpidkhud thkj u~cudadl ~hmh ucjick eck}pho+ Pchlud ^EPH mhk @ mdbhmh~lhk ~hmh ucjick gqcpoh~~dkj thkj mdhi~odadlhud ~hmh MKH thkj
ickjhkmskj is}hud thkj }cohb mdihusllhk "mckjhk ici~cp}di`hkjlhk iduih}ebdkj
`huc ickjjskhlhk ~pdicp is}hk} 8I h}hs 0I$+ Uc}cohb msh ~pgmsl mdlgi dkhudlhk/
mckh}sphud mhk pchkkcho mdohlslhk/ ~godicphud MKH mh~h} ici~cp~hkfhkj sfskj 5
bc}cpgms~ocl mckjhk sfskj 5 thkj }cpuci`sktd+ Lcismdhk/ ~pgmsl ~hkfhkj ~cksb
mckjhk is}hud thkj mdihusllhk ~hmh ucjick eck}pho mh~h} mdhi~odadlhud mckjhk
ickjjskhlhk bhkth ~pdicp gs}cp 8 mhk 0+
) ^EP @huc ]skjjho Iduih}ebcm
^hmh hzho ~ckjci`hkjhk ic}gmc ^EP/ hi~odadlhud MKH ickskfsllhk `hbzh }clkdl @huc
}skjjho iduih}ebcm uhkjh} ~g}ckudho sk}sl mdh~odlhudlhk ~hmh is}hjckcudu "Uebhp} c} ho+
8421$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 58
-
7/23/2019 Protein Engineering _ Siti Julaiha
32/98
Udkjoc `huc iduih}ebcm hk}hph hi~odadlhud ~pdicp mhk }ci~oh}c mdlgp~gphudlhk ~hmh
ususkhk }ci~oh}c uc`hjhd bhudo hi~odadlhud+
Bd}sebd c} ho+ "8422$ ickcphkjlhk `hbzh qhpdhud ic}gmc mhuhp iciskjldklhk is}hud ~hmh
~pgmslud aphjick MKH mckjhk ^EP+
Msh ~pdicp pchlud ^EP ici~pgmslud pchlud mckjhk icohlslhk }si~hkj }dkmdb h}hs
gqcpoh~~dkj msh aphjick MKH/ thkj ucishkth ickhi~skj is}hud thkj uhih ~hmh
mhcphb gqcpoh~kth+
Ususkhk thkj gqcpoh~ iciskjldklhk }cp`ck}slhkth aphjick ickfhmd bd`pdmduhud+ Uhohb
uh}s mhpd msh bt pdm md~cp~hkfhkj mckjhk MKH ~oodicphuc sk}sl ici~pgmslud aphjick
ms~oclu+ Bd`pdm uh}skth icidodld pcecuucm sfskj 3 mhkl hpckh `slhk uc`hjhd us`u}ph}c
sk}sl ~godicphud/ ihlh hlhk ucehph cacl}da bdohkj mhpd ~cpehi~sphk pchlud+ Uc~cp}d ~hmh
is}hjckcdud lgkqcpkudgkho ~cp~hkfhkjhk ~pdicp/ mhk is}hud ~ckjbdohkjhk mhk h}hs
~ckhi`hbhk mhcphb huhi hidkg mh~h} fsjh mded~}hlhk+
Ohkjlhb thkj md}skfsllhk ~hmh `hjdhk ldpd h}hu sfskj mdhjphi ickskfsllhk ic}gmc `hude
^EP is}hjckcudu+ Uc}ckjhb `hjdhk lc `hzhb ickskfsllhk `hjhdihkh is}hud mh~h}
md~dkmhlhk ~hmh igoclso MKH `hjdhk }ckjb+ ^pdicp md}skfsllhk mckjhk bspsa }c`ho mhk
~pdicp H mhk H uhodkj icockjlh~d+
Ic}gmc Bdjsebd "8422$ ucmdld} psid} lhpckh ici`s}sblhk ci~h} ~pdicp mhk 5 ^EPu
"uc~huhkj ^EPu sk}sl hi~odadlhud ucjick gqcpoh~~dkj mhk }djh ^EP sk}sl
ickjjh`skjlhk msh ucjick$+
Pchjck Lgicpudho ~pgmsuck }cohb ickjci`hkjlhk `c`cph~h ic}gmc thkj oc`db
ucmcpbhkh mhk msh mhpd ic}gmc dkd mdmculpd~udlhk md`hzhb 6
8+ ^pgucmsp thkj }dmhlici`h}hud ~cps`hbhk `huc }skjjho/ mckjhk icidodb ~pdicp thkj
ucushd sk}sl md}hi`hblhk h}hs mdbdohkjlhk+
0+ ]hx ~godicphuc thkj ickthodk MKH mckjhk admcod}hu pckmhb/ mckjhk pcudlg is}hud thkj
clu}phkcgsu }cpfhmd ucohih pchlud hi~odadlhud+ ^pg oci dkd mdlsphkjd mckjhk
ickjjskhlhk ~godicphud }cpigu}h`do mckjhk admcod}hu }dkjjd/ mhk lgkuck}phud }ci~oh}cthkj }dkjjd ucp}h ~ckjsphkjhk udlosu hi~odadlhud mhpd 8)udlosu+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 50
-
7/23/2019 Protein Engineering _ Siti Julaiha
33/98
) ^EP Ic}gmc Crud}c ]I+
^hmh ic}gmc Crud}c ]I/ lcmsh phk}hd qcl}gp thkj ici`hzh }hpjc} jck thkj hlhk
mdhi~odadlhud ickjjskhlhk ^EP }c}h~d uh}s mhpd ~pdicp ici`hzh is}hud thkj mddkjdklhk+
Bhudo ~pgmslud mhpd ~g~sohud odkcp ms~ocr ici`hzh jck is}hud thkj hlhk mdlgk}hidkhudlhk
mckjhk }dkjlh} pckmhb ~hmh udlosu }ci~oh}c MKH huodkth+
]ci~oh}c MKH md}spsklhk mhpd Uco C)egod mckjhk udu}ci igmdadlhud pcu}pdlud dk}hl / lcismdhk
mdohlslhk ic}dohud mhk mduckud}dalhk sk}sl pcu}pdlud mckjhk M~kDCkmgkslochuc+
MKH odkchp md~pgmslud mckjhk hi~odadlhud hlhk pcudu}hk }cpbhmh~ ~ci`slhhk M~ko mhk
uc}cohb udplsohud mckjhk odjhud `osk})ckm mh~h}d mdlci`hodlhk mckjhk }phkuagpihud lcmhohi
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 55
-
7/23/2019 Protein Engineering _ Siti Julaiha
34/98
C)egod+ Uc}dh~ igoclso bd`pdm ickjhkmskj phk}hd }skjjho }ci~oh}c ic}dohud MKH mhk
skic}dohud hi~odadlhud thkj hlhk mdbhkesplhk mckjhk Ckmgkslochuc+
) ^EP Ic}gmc Jck ]hdogp
Ic}gmc dkd ickhpjc}lhk MKH thkj mdic}dohud dk qd}pg uc`cosi ohkjlhb is}hjckcudu mhk
gqcpoh~~dkj ~pdicp mdjskhlhk+
Odkdcp hi~odadlhk thkj md~pgmslud/ ici`hzh is}hud thkj mddkjdklhk }c}h~d uhh}
md}phkuagpihudlhk ohkjuskj ~hmh C)egod/ Uco Bgu} ici~cp`hdld udplsohud ckvdi is}hud
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 5=
^EP Ic}gmc Crud}c ]I
^EP Ic}gmc Jck
-
7/23/2019 Protein Engineering _ Siti Julaiha
35/98
MKH odkchp/ ucick}hph Ckmgkslochuc [email protected] ickjsphdlhk Ic}doh})}ci~h}c MKH/
ickdkjjholhk bhkth sk)ic}doh} uc`hjhd ~pgmsl is}hud+
) Pcqcpuc ]phkulpd~ud ^EP
Ic}gmc uckud}da sk}sl ickmc}clud mhk ickjhkhoduh }phkulpd~ud iPKH h}hs PKH ~hmh ~gudud
thkj pckmhb+ PKH }dmhl mh~h} mdjskhlhk uc`hjhd }ci~oh}c ^EP ucbdkjjh bhpsu
md}phkulpd~udlhk mhbsos ickfhmd eMKH mckjhk pcqcpuc }phkulpd~ud mhk Igogkct qdpsu h}hs
Hqdhk qdpsu mhk ms~odlh} eMKH lcismdhk md~cp`hkthl+ ]clkdl dkd mdhzhod mckjhk
~ckehi~sphk idrdkj ubgp} ~godicp }didmdkc "godjgm]$ mckjhk iPKH mhk hkkcho PKHu
~godhmckhod} }hdo+ Pcqcpuc }phkulpd~ud md}hi`hblhk mhk mdjskhlhk godjg m] uc`hjhd ~pdicp
sk}sl udk}cuh ~cp}hih phk}hd eMKH+
) ^EP ) Jck Uclsckud sk}sl Is}hud
^pgmsl ^EP sk}sl Is}hud jck thkj mduhpdkj mdmh~h}lhk mhpd Jckgic MKH ickjjskhlhk
~pdicp dk}pgk)u~cuadl thkj ickjh~d} crgk h}hs mckjhk Pcqcpuc ]phkulpd~ud EP+
"H$ Jckgic MKH/ Crgku 8)= mh~h} mdhi~odadlhud ucehph }cp~duhb mhpd Jckgic MKH
ickjjskhlhk ~huhkjhk ~pdicp dk}pgk thkj u~cudadl "8A , 8P/ 0A , 0P/ mu`+$
"@$ Pcqcpuc ]phkulpd~ud ^EP+ Ici~cpehthlhk ~hmh `c`cph~h iPKH thkj
md~pcuck}hudlhk ~hmh uco uco thkj ucehph ismhb mdhlucu uc~cp}d uco uco mhphb/
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 53
-
7/23/2019 Protein Engineering _ Siti Julaiha
36/98
iciskjldklhk lgkqcpud lc eMKH+ eMKH lcismdhk mh~h} mdjskhlhk uc`hjhd }ci~oh}c
sk}sl ~huhkjhk ~pdicp crgk thkj u~cudal sk}sl icked~}hlhk Aphjick MKH thkj
gqcpoh~~dkj'}si~hkj }dkmdb+
) ^EP) ^cp~hkfhkjhk ^godigp~b Aphjick Pcu}pdlud Aphjick}
Ic}gmc sk}sl ickmc}clud mhcphb pcu}pdlud ~godigp~+ Hoococu 8 mhk 0 md`cmhlhk mckjhk
~godigp~b thkj icps`hb ususkhk kslocg}dmh mhpd mhcphb pcul}pdlud u~cudadl sk}sl pcu}pdlud
kslochuc+ Hoococ 8 ici~skthd mhcphb/ }c}h~d hocooc 0 ici~skthd kslocg}dmh thkj }cohb
mdps`hb+ ^EP ~pdicp mh~h} mdmcuhdk ucehph ucmcpbhkh mhpd ususkhk thkj ickjh~d} mhcphb
pcu}pdlud sk}sl ici~pgmslud ~pgmsl thkj ~ckmcl+ ^ckjbhkesphk ^pgmsl ^EP mckjhk
ckvdic P mhk aphludkhud slsphk mh~h} ickjbhudolhk ~cksoduhk ucmcpbhkh sk}sl msh hocoocu+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 51
-
7/23/2019 Protein Engineering _ Siti Julaiha
37/98
) ^EP U]P^U
^EP mh~h} mdjskhlhk sk}sl ickjbhudolhk }hkmci ~ckmcl ~godigp~u thkj `cpsohkj
"U]P^u$+ ^cksoduhk mhpd mdkslocg}dmh EH']J ickjsohkj ~godigp~bu thkj ici~skthd }djh
hoco uc`hjhd bhudo qhpdhud ~hmh fsiohb ~ckjsohkjhk EH+ ^hmh hs}gphmdgjph~b uc}dh~ hoco
mdpc~pcuck}hudlhk mckjhk `hkm s}hih `hjdhk h}hu mhk msh idkgp `hkm `hthkjhk+ Uc}dh~
H mhk @ ici~skthd jckg}d~c H "8/5$ mhk @ "0/0$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 5:
-
7/23/2019 Protein Engineering _ Siti Julaiha
38/98
) ^EP U~cudadl Hoco
Mhuhp mhpd ^EP u~cuadl hoco dkd ickjlgpclud ~huhkjhk `huc ~hmh sfskj 5 ^pdicp ^EP+
U~cudl hoco godjgkslocg}dmh ~pdicp HU^8 mhk HU^0 mdphkehkj sk}sl ickthihlhk ususkhk
msh hoco }cpbhmh~ ~gudud mhcphb uc`cosikth ~hmh qhpdhk kslocg}dmh/ uhi~hd mckjhk mhk
`cphlbdp ~hmh qhpdhk d}s uckmdpd+ HU^8 hlhk ickjdlh} ucehph uci~spkh sk}sl icockjlh~d
ususkhk phk}hd hoco 8/ iciskjldklhk hi~odadlhud mckjhk ~pdicp thkj mdodkmskjd+ Khisk/sfskj E)5 ~hmh HU^0 ~pdicp iduih}ebcm mckjhk ] mhpd ususkhk hoco 8/ ucbdkjjh
hi~odadlhud }dmhl mh~h} mdohlslhk+ Ucps~h mckjhk HU^0 mh~h} ickjdlh} uci~spkh lc~hmh
hoco 0 mhk icisohd hi~odadlhud/ }dmhl uc~cp}d HU^8+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 52
-
7/23/2019 Protein Engineering _ Siti Julaiha
39/98
)^EP MG^
Iciskjldklhk logkdkj eMKH ickjjskhlhk }spshk godjgkslocg}dmh+
Logkdkj eMKH sk}sl ~gpedkc sph}c gludmhuc ikcjjskhlhk }spskhk godjgkslocg}dmh thkj
`cpbs`skjhk mckjhk ususkhk huhi hidkg thkj mdlc}hbsd+ ^pdicp uckuc mdlgku}psludlhk
}cpbhmh~ lgmgk :)88 mhk msh `huc thkj ~cp}hih mhpd lgmgk 80/ mhk ~pdicp hk}duckuc
`cpbs`skjhk mhkjhk lgmgk 5=)52 "Occ c}+ho/ 842>$+ Ususkhk huhi hidkg md~dodb sk}sl
ickjlgku}psludlhk ~pdicp thkj md~dodb mhpd `hudu lhkmskjhk }dkjjd huhi hidkg thkj
mdu~cudadlhk mckjhk bhkth msh lgmgk "Hu~/ ]tp/ Otu/ Huk/ Bdu$+ ^pdicp ici~skth~cp~hkfhkjhk 3 ickjhkmskj ususkhk thkj mdlckhod sk}sl pcu}pdlud kslochucu/ sk}sl
iciahudod}hud logkdkj us`uclsck uco `huc+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 54
-
7/23/2019 Protein Engineering _ Siti Julaiha
40/98
) ^EP Odklcm ^pdicp
^EP odkl ~pdicp iciskjldklhk hi~odadlhud dk)mdulpdidkh}c ususkhk MKH ~hmh }hpjc} MKH
thkj psid}'lgi~oclu+
Igoclso Odklcm "hmh~}gp$ hmhohb phk}hd jhkmh godjgkslocg}dmh thkj md`ck}sl mckjhk odjhud
msh phk}hd }skjjho godjgkslocg}dmh mdockjlh~d ~hmh ususkhk lceshod thkj icidodld
lgi~h}d oc }cpjhk}skj 3 mckjhk pc}pdlud kslochuc }cpjhk}skj'gqcpbhkj "~hmh lhusu dkd/
JH]E 3 }cpjhk}skj md~pgmslud gocb I`go$+ Uc}cohb odjhud mhpd odklcp }cpbhmh~ }hpjc}
aphjick pcu}pdlud/ ~pdicp odklcm)u~cudadl mh~h} ickjbhudolhk hi~odadlhud ucish aphjick
mckjhk ickjdlh} msh igoclso odklcp thkj ickjh~d}+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =>
-
7/23/2019 Protein Engineering _ Siti Julaiha
41/98
) Hkebgpcm ^EP Jckgic Zholdkj
Usi`cp lgi~ocl }hpjc} mhpd MKH thkj }cpmdpd mhpd aphjick thkj `hkthl mdihkh ususkhk
hkebgp "oh`sbhk$ mdohi~dplhk/ uc`hjhd egk}gb odklcp godjgkslocg}dmh phk}hd jhkmh+ Dmc sk}sl
ickjjskhlhk u~cudadl ~pdicp sk}sl ususkhk hkebgp mhk uh}s u~cudpdl sk}sl ususkhk R
thkj mdlc}hbsd mh~h} sk}sl ici`c`hulhk aphjick aphjic thkj ickjhkmskj ususkhk R
mhk ickmh~h}lhk hlucu ususkhk }dmhl }cpdmck}dadlhud uc`cosikth thkj `cpmhi~dkjhk
}cpbhmh~ R+ ^hmh egk}gb dkd/ ususkhk hkebgpcm md}skfsllhk bhkth ~hmh `hjdhk udud ldpd mhpd
hi~odadlhud thkj fcohu mhk mdiskjldklhk mhpd ususkhk lhphl}cpduhud K8 uc`cosikth thkj
`cpmclh}hk mckjhk ususkhk R thkj mdlc}hbsd+ Qhpdhud ic}gmc mcpdqh}da }cohb mdususk/
uc~cp}d EP s`oc)odklcp+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =8
-
7/23/2019 Protein Engineering _ Siti Julaiha
42/98
Hkebgpcm EP Jckgic Zholdkj ^EP ) ]hxihk Huuht
) ^EP ) ]hxihk Huuht
Ic}gmc uckud}da sk}sl ickmc}clud u~cudadl Hocooc ickjjskhlhk hl}dahud crg 3 mhk 5 mhk
~pdicp lc}djh mckjhk Aosgp mhk Xsckebcp+
^ckhi`hbhk 0 ~pdicp ^EP lgkqckudgkho/ ^8 mhk ^0/ thkj u~cudadl sk}sl ususkhk }hpjc}/
~pdicp lc}djh ^5/ mdmcuhdk sk}sl ickjdlh} ucehph u~cudadl mhcphb ~hmh ususkhk }hpjc}
mgzku}pchi ~hmh ush}s dlh}hk ^8+ ^coh`cohk ^5 mckjhk msh aosgpg~bgpcu/+ pc~gp}cp mtc
"P$ mdohi~dplhk ~hmh sfskj 3 mhk xsckebcp mtc thkj ici~skthd ~cp`cmhhk ~hkfhkj
jcogi`hkj cidud mdohi~dplhk ~hmh pc~gp}cp mtc thd}s ~hmh sfskj 5+ Lhpckh sfskj 5
md`ogl'mdbhohkjdk/ ^5 ~pdicp lc}djh }dmhl mh~h} mckjhk uckmdpdkth ~pdic ~hmh uc}dh~ udk}cuh
MKH `hps+ Ucohih pchlud ^EP/ ^godicphud ]hx MKH ickudk}cuh phk}hd ~pdih MKH `hps
mckjhk ^8 ' ~pdicp ~cp}hih mhk ckvdi ickmclh}d ~pdicp lc}djh/ hl}dad}hu ~pgucu mhpd hphb 3
5 ickspsklhk ^5 ~pdicp lc}djh mhpd sfskj 3+ Bhudo hlbdp hmhohb phk}hd MKH thkj }di`so
icoc`dbd mhcphb dlh}hk ^5 pc~gp}cp ucp}h xsckebc mtc }dmhl ohjd ickjdlh} igoclso thjk
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =0
-
7/23/2019 Protein Engineering _ Siti Julaiha
43/98
uhih+ Lhpckh Pc~gp}cp mtc }dmhl ohjd mclh} ~hmh xsckebcp/ bhudokth ickdkjlh}lhk cidud
dk}ckud}hu pc~gp}cp thkj hlhk mckjhk ismhb mdmc}clud
) ^EP ) Bg} U}hp
]clkdl thkj ickjs`hb ehph ~ckehi~sphk ^EP mckjhk ~cihkhuhk hzho+ ^godicphuc
mdhl}dalhk/ }h~d }hpjc} `cosi ickjhohid mdmckh}sphud mhk ^pdicp mh~h} ickjdlh} lc oglhud
kgk)u~cudadl "h}hs `hblhk uh}s uhih ohdkkth$+ ]clkdl mdohlslhk ihksho mckjhk
icihkhulhk lgi~gkck pchlud ~hmh usbs ~ckehdphk "idu+ 43 E$ uc`cosi ickhi`hblhk
~godicphuc + H}hs/ udu}ci thkj ickjbhi`h} hl}dqd}hu ~godicphuc thkj ~hmh usbs
odkjlskjhk/ `hdl gocb ~ckjdlh}hk ush}s hk}d`gmd/ h}hs mckjhk hmhkth dkbd`d}gp thkj }cpdlh}
lgqhock thkj bhkth }cpmdugudhud uc}cohb ohkjlhb hl}dqhud }ci~cph}sp }dkjjd+ Bg})u}hp}'egom)
adkdub ^EP mdeh~hd mckjhk ~godicphuc bd`pdmh `hps thkj }dmhl hl}da ~hmh usbs odkjlskjhk
mhk bhkth mdhl}dalhk ~hmh }ci~cph}sp }dkjjd+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =5
-
7/23/2019 Protein Engineering _ Siti Julaiha
44/98
^godicphuc ]hx `cphuho mhpd `hl}cpd ]+ hxsh}desu mddmck}dadlhud uc`hjhd ckvdi thkj ihi~s
ickhbhk lgkmdud ~pg}cdk thkj md~duhblhk gocb usbs }dkjjd ucohih ~pgucu ^EP+
Usbs ]hx*u g~}diho sk}sl lcjdh}hk hmhohb :3)2> E/ mckjhk zhl}s ~hpsb mhpd 4 ickd} ~hmh
4:/3 E/ mhk mh~h} ickdps 8>>> sk}hd ~huhkjhk `huh MKH mhohi zhl}s lsphkj mhpd 8>
mc}dl ~hmh :0 E+
]hx MKH ~goticphuc ohelu }bc 53 ~pgga)pchmdkj he}dqd}t thkj `dhuh }cpmh~h} ~hmh
~goticphucu+ ]hx idu)dkegp~gph}cu 8 `huc dk 8>=+
H =>> `~ }hpjc} hlhk ickjhkmskj cppgp 55# ~hmh igocesocu uc}cohb 0> eteocu+
Cppgp mdu}pd`s}dgk zdoo `c phkmgi+ @dhuhkth icidodld }dkjlh} lcuhohbhk mdslsp ucld}hp 8 mhohi
4+>>> kslocg}dmh Cppgp md~cpbh}dlhk `doh }hpjc} dkjdk ici~cp`hkthl MKH dkmdqdmsho
igocesoc mh~h} icktci`sktdlhk `c`cph~h is}hud+ Cppgp }dmhl md~cpbh}dlhk fdlh }hpjc}
ickjdkjdklhk ususkhk hi~odadcm MKH h}hs icig}gkjkth mckjhk pcu}pde}dgk ckvticu+
^pgga)pchmdkj }bcpig)u}h`oc ckvtic uc~cp}d MKH ~godicphuc ^as/ icidodld hl}dqd}hu
~pggapchmdkj/ mhk mdjskhlhk uc`hjhd ~ckjjhk}d "h}hs mhohi lgi`dkhud mckjhk$ ]hx sk}sl
hi~odadlhud lchlsph}hk thkj }dkjjd +
Hi~odadlhud ~pgmsl `dhuhkth ~hmh phkjc udvc 8>>)83>> `~+
]hpjc} thkj oc`db ~hkfhkj md ~cp`hkthl ;03 l`+
Ici`s}sblhk igmdadlhud ~hmh pchlud `saacp/ gel}hdou ~goticphucu/ mhk zhl}s clu}ckud thkj
oc`db ohih+
) Cppgp ^pgkc ^EP
]clkdl ^EP mckjhk lc}cod}dhk thkj pckmhb+ Mdjskhlhk sk}sl ickck}slhk qhpdhku `hps
~pg}cdk `cpmhuhplhk ~pdkud~ `hbzh ]hx ~godicphuc mh~h} `cphkkchodkj ~hmh ~huhkjhk
`huc thkj }dmhl lgi~h}d`oc uh}s uhih ohdkkth ucohih ~pgucu hi~odadlhud h}hs ~ckjsh}hk
mhohi lgkmdud ^EP thkj }dmhl uci~spkh+
]clkdl dkd mddohlslhk mckjhk ickdkjlh}lhk lgkuck}phud dgk Ij 0, / ~ckhi`hbhk dgk Ik 0,/
Ici`cmhlhk Lgkuck}phud ~hmh lcci~h} mK]^u/ ^ckjjskhhk mD]^/ mhk Ickdkjlh}lhk
fsiohb ]hx ^godicphuc "^godicphuc }hk~h `sl}s askjud ~ci`hehhk$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc ==
-
7/23/2019 Protein Engineering _ Siti Julaiha
45/98
Mhk `hkthl ohjd egk}gb ]clkdl }clkdl ^EP thkj mdjskhlhk mhohi pclhthuh Jckc}dl+
U]PH]CJD MCUHDK IS]HJCKCUDU HEHL' PHKMGI
IS]HJCKCUDU
Bhudo ~cpeg`hhk }clkdl lhmhkjlhoh ici`cpdlhk ~cps`hbhk thkj mddkjdklhk mckjhk hmhkth
is}hud thkj }dmhl md~cpldphlhk+
Mhohi }clkdl dkd }dmhl md~cposlhk ~ckjc}hbshk }ck}hkj u}psl}sp h}hs ususkhk ~pg}cdk/ h}hs~sk
iclhkduic ~pg}cdk/ icohdklhk ~ckcph~hk ~ckjc}hbshk mhohi 6
) Phkmgi is}hjckcudu
Mdjskhlhk sk}sl icked~}hlhk ~cp`cmhhk jckc}dl
) Ucoclud h}hs Uepcckdkj'^ckthpdkjhk
Mdjskhlhk sk}sl icidodb qhpdhk thkj icidodld lshod}hu thkj mddkjdklhk+
Mhohi `c`cph~h }hbh~hk is}hud mhk ucoculd/ ic}gmc dkd ickdps cqgosud hohi+ @dhuhkth
mdjskhlhk sk}sl ickjbdohkjlhk is}hud kc}pho mhk icpsuhl+
^pdkud~ mhuhp thkj ickmhuhpd mcuhdk u}ph}cjd dkd hmhohb sk}sl iciqhpdhudlhk huhi hidkg
ucehph phkmgi h}hs hehl ~hmh ususkhk ~god~c~}dmh sk}sl ickmh~h}lhk lgi`dkhud lgi`dkhud
`hps mhk lcismdhk icktcoclud ~pgmsl ~pgmsl jck `hps "~pg}cdk$ +
Egk}gb is}hjckcudud hehl'phkmgi thkj mdjskhlhk mhohi pclhthuh ~pg}cdk hmhohb Cqgosud
thkj mdhphblhk "Mdpce}cm cqgosud$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =3
-
7/23/2019 Protein Engineering _ Siti Julaiha
46/98
]clkdl ~ckmclh}hk mckjhk cqgosud thkj mdhphblhk iciskjldklhk sk}sl icked~h}lhk ~pg}cdk
~pg}cdk `hps mhk ~pgmsl ~pgmsl jck mckjhk is}hud thkj mddkjdklhk ickjjskhlhk }clkudl
is}hjckcudu es}}dkj)cmjc "~cig}gkjhk sfskj$ thkj ickfhmd }clkdl ~cphkehkjhk lci`hod
~pg}cdk ~pg}cdk thkj }cohb cludu h}hs hmh+ Khisk/ ucehph cadudckud `hbhk ldidh lcucospsbhk
mhpd ~cphkehkjhk lci`hod ~pg}cdk ~pg}cdk mh~h} ickjsphkjd kvdi thkj }cpfhmd ucehph kh}sppho+
Uhkjh} mddkjdklhk h}hs md~dodb gocb ~hph ~ckcod}d sk}sl ickhi~dolhk huuht cqgosud mdpcl dkd ~hmh
is}hud is}hud thkj ickjbhudolhk lgkmdud u}h`do }cpbhmh~ }clhkhk }clhkhk uc~cp}d }ci~cph}sp/
~B moo / md}ci~h}lhk ~hmh lso}sp ~g~sohud uco ickjjskhlhk dku}psick} uc~cp}d lcigu}h}+
Mckjhk iskesokth }clkdl sk}sl icohlslhk is}hjckcudu hehl'phkmgi dkd iciskjldklhk sk}sl
ickfcohulhk bs`skjhk u}psl}sp ~pg}cdk)askjud mckjhk ici~cp`hdld h}hs ickjs`hb sps}hk
~god~c~}dmh+ Is}hud dkd md~cplckholhk mckjhk ickjjskhlhk ic}gmc thkj mduc`s} Cppgp ~pgkc
^EP "^goticphuc Ebhdk Pche}dgk$+ ^godicphuc thkj mdjskhlhk mhohi ^EP pck}hk }cpbhmh~
lcuhohbhk/ ucbdkjjh }di`so lcuhohbhk mhohi ici~cplckholhk sps}hk jck+ ^pgmsl is}hud
^EP dkd lcismdhk mdodjhudlhk lc mhohi ~ohuidm clu~pcud ~pg}cdk mhk ~cp~su}hlhhk is}hk
mh~h} mduhpdkj sk}sl is}hud ~hmh ~pgmsl jck h}hs ~pg}cdk+
]hbh~hk mhohi Phkmgi Is}hud hmhohb 6
8+ Icidodb Jckc
0+ Icked~}hlhk od`phpd'~su}hlh mhpd qhpdhk qhpdhk
5+ Icihusllhk ~su}hlh jck lc~hmh qce}gp thkj mdclu~pcudlhk
=+ Icihusllhk ~su}hlh jck h}hs qce}gp lc `hl}cpd/ thkj ici~pgmslud qhpdhk qhpdhk
ckvdi mhpd uh}s uco h}hs uh}s ususukhk
3+ Ickthpdkj lgogkd lgogkd sk}sl udah} udah} thkj mddkjdklhk
1+ Ickjdugohud jck jck thkj hlhk mdlci`hkjlhk mhk ickjsohkjd ~pgucu
Uc~cp}d thkj }cpjhi`hplhk cpdls} dkd 6
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =1
-
7/23/2019 Protein Engineering _ Siti Julaiha
47/98
Ahl}gp thkj ickck}slhk lc`cpbhudohk u}ph}cjd hmhohb6
Lshod}hu mhk mcuhdk ^su}hlh'Od`phpt
^cidodbhk ic}gmc cqgosud
^cidodbhk Pclgi`dkhud MKH
Ic}gmc Ucoclud h}hs ^ckthpdkjhk'Ulpcckdkj
Odid}hud h}hs lc}cp`h}huhkthkj mdbhudolhk mhpd u}ph}cjd dkd hmhohb6
Ususkhk ~pg}cdk thkj mdlci`hkjlhk icidodld udah} thkj bhi~dp idpd~ mckjhk
ususkhk dkmsl h}hs ~hpck}ho/ uc~hkfhkj ~h}b cqgosudkth+
Lh~hud}hu ic}gmc sk}sl icked~}hlhk ususkhk askjudgkho thkj `hps uhkjh}
}cp`h}hu mhk ici`s}slhk od`phpt'~su}hlh thkj `cuhp+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =:
-
7/23/2019 Protein Engineering _ Siti Julaiha
48/98
CQGOSUD THKJ MDHPHBLHK'MDPCE]CM CQGOSUD
Sk}sl icihbhid cqgosud mdhphblhk/ ihlh md~cposlhk ~cihbhihk }ck}hkj cqgosud+ Ucoclud
hohi/ `cpuhih mckjhk lgi`dkhud jck)~pg}cdk/ `cp dk}cphlud/ ickmgpgkj cqgosud+ Mhohi
cqgosud/ askjud ckvdi }cp}ck}s mhk udah} ckvdi cps`hb/ icktci~spkhlhk mhk mducushdlhk
mckjhk gpjhkduic odkjlskjhk+ ^cps`hbhk dkd }dmhl `clcpfh mhohi hphb }cp}ck}s h}hs }cpbhmh~
h~h~sk }sfshk thkj }c~h}/ }c}h~d }cpfhmd ucehph u~gk}hk ~hmh `ck}sl thkj uhkjh} u~cudho mhohi
ickhkjjh~d pc~pgmslud u~cudal mhk ~cputhph}hk bdms~+ Ld}h }hbs dkd/ }h~d ld}h }dmhl }hbs
`hjhdihkh }c~h}kth dk}cphlud ~pgucu `clcpfh+ Fsjh bho h~h thkj `clcpfh mckjhk `hdl mhohi
odkjlskjhk ucosohp }dmhl cacl}da mhohi odkjlskjhk kgkkh}spho sk}sl `c`cph~h hohuhk/ uc~cp}d
us`u}ph}c thkj }ml m~} icohps}/ ~pgmsl thkj }dmhl u}h`do h}hs pchlud ldidh }hi`hbhk++ Ucbdkjjh
ckvdi hohi thkj mdjskhlhk mhohi ~cp}hkdhk/ lcmgl}cphk mhk dkmsu}pd h~odlhud iskjldk }dmhl
ucohos ici`cpdlhk bhudo thkj mh~h} mdhkmholhk+ H~h thkj md`s}sblhk hmhohb ckvdi)ckvdi
thkj u}h`do thkj mh~h} }c}h~ hl}da mhohi fhkjlh ~cpdgmc zhl}s thkj ohih ~hmh odkjlskjhk kgk)
ucosohp mhk thkj mh~h} `clcpfh mckjhk `cp`hjhd us`u}ph}+ Md udkdohb cqgosud thkj mdhphblhk
mh~h} mdh~odlhudlhk+
Cqgosud thkj mdhphblhk icps~hlhk ~pgucu thkj mdlckmhodlhk mhohi `dgogjd udk}c}du thkj
mdjskhlhk sk}sl icpclhth ~pg}cdk h}hs PKH mckjhk udah})udah} }cp}ck}s thkj mddkjdklhk thkj
iskjldk h}hs }dmhl iskjldk }cpfhmd ucehph hohid+ Mhohi fckdu pclhthuh ~pg}cdk dkd/ doiszhk
`clcpfh icksfs }sfshk thkj mdmcadkdudlhk ucehph ~hu}d ) iskjldk uc`ckhpkth hmh `hkthl ehph
sk}sl ickeh~hd }sfshk ~hu}d/ khisk ~ckcod}d icidodb uhohb uh}s thkj ~hodkj }dmhl icicposlhk
s~hth ) mhk icigkd}gp lcucospsbhk ~pgucu+
Clu~cpdick Mdpce}cm Cqgos}dgk
Clu~cpdick cqgosud thkj mdhphblhk mh~h} mdohlslhk ~hmh uco)uco bdms~ "dk qdqg$ mhk fsjh }hk~h
ickjjskhlhk uco bdms~ "dk qd}pg$+ Uhohb uh}s ~s}hphk ~cpeg`hhk `dhuhkth ickjhi`do }djh
ohkjlhb sk}sl icktcocuhdlhk mhk ~s}hphk `c`cph~h ~cpeg`hhk `dhuhkth md~cposlhk sk}sl
ickmh~h}lhk ~pg}cdk h}hs PKH mckjhk edpd)edpd thkj md~cposlhk+
Sk}sl icisohdkth/ mdohlslhk ohkjlhb ickjdugohud jck)jck mckjhk udah})udah} u~cudadl thkj
mddkjdklhk mhk/ mckjhk ickjjskhlhk ~pgucu is}hjckcudu mhk ' h}hs pclgi`dkhud/ ici`sh}
~cp~su}hlhhk cuhp qhpdhk jck< ucihldk cuhp ~cp~su}hlhhk ucihldk cuhp lcuci~h}hk sk}sl
ickcislhk qhpdhk jck mckjhk udah})udah} thkj md`s}sblhk+ Lcismdhk ~su}hlh qhpdhk jck
mduhpdkj sk}sl ickcislhk mhk ickjdugohud qhpdhk jck thkj md}dkjlh}lhk mhk lcismdhk
mdhi~odadlhud `cplhod)lhod mhk mduhfdlhk mhohi `hl}cpd/ fhisp h}hs gpjhkduic ohdk thkj egegl+
Mhohi ~s}hphk ~cpeg`hhk cpdls}kth/ qhpdhk jck }cp~dodb mhpd ~s}hphk uc`cosikth ucohkfs}kth
md}dkjlh}lhk ohjd mhk `cjd}s uc}cpsukth uhi~hd hlbdpkth mdmh~h}lhk jck mckjhk udah})udah} thkj
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =2
-
7/23/2019 Protein Engineering _ Siti Julaiha
49/98
md`s}sblhk+
IC]GMC KGK [email protected] MGIHDK UZH^^DKJ
Lgku}pslud hl}sho mhpd mcuhdk phudgk jckcu mgihdk uzh~~cm jckcu mh~h} mducocuhdlhk
mckjhk c`cph~h }clkdl thd}s 6
) Ic}gmc Lhuc} Is}hjckcudu
Ususkhk ud}c pcu}pdlud ickjh~d} ususkhk MKH }cp}ck}s sk}sl md}ci~h}lhk mhk
mdbhkesplhk mckjhk ckvdi pcu}pdlud mhk ~ckjjhk}dhk ususkhk mckjhk
icihusllhkkth md hk}hph ud}cukth "Zcoou c}+ho+8423$+
) Ic}gmc ^cig}gkjhk Jckc'U~odedkj mckjhk ^EP ~cp~hkfhkjhk gqcpoh~~dkj
Igoclso MKH mdpclgi dkhudlhk }hk~h ickjjskhlhk ~ci`h}huhk ckmgkslcohuc
"Bgp}gk/ c}+ho 8424$+
Aphjcick jck thkj hlhk mdpclgi`hkdulhk mhpd jck gphkj}sh mh~h} mdbhudolhk
mhohi pchud ^EP+
IC]GMC LHUC] IS]HJCKCUDU
^ci`h}huhk ud}su mh~h} }cpfhmd ucehph hohid h}hs hp}dadudho mdisohd ~hmh oglhud thkj mddkjdklhk
~hmh jck }hpjc} mckjhk godjgkslocg}dmh)mdpce}cm is}hjckcudu+
^cidodbhk ~ci`h}huhk ud}su mdmhuhplhk ~hmh lcskdlhkkth }cpbhmh~ ~ohuidm mhk lgkucpqhud
huhi hidkg hlbdp sps}hk ~ckjlgmchk+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =4
-
7/23/2019 Protein Engineering _ Siti Julaiha
50/98
IC]GMC ^CIG]GKJHK JCK MCKJHK ^EP GQCPOH^ CR]CKUDGK
^pdicp sk}sl pchlud)pchlud dkd mdmcuhdk ucbdkjjh sfskj ~pgmsl ickjhkmskj sps}hk
lgi~ocick}cp+
Lc}dlh ~pgmsl ^EP dkd mdehi~sp/ hosp icidodld sps}hk thkj ucushd ~hmh sfskj phk}hd 5 thkj
}si~hkj }dkmdb mhk `cp}dkmhl uc`hjhd ~pdicp uh}s uhih ohdk/ mhk clu}ckud gocb ~godicphuc MKH
ickjbhudolhk ush}s sps}hk md ihkh sps}hk huod mdbs`skjlhk `cpuhih)uhih+
Iciskjldklhk sk}sl ickjjh`skjlhk msh aphjick mhohi uh}s ohkjlhb logkdkj/ khisk
ohkjlhb logkdkj iso}d~oc md`s}sblhk sk}sl icohlslhk uzh~ mgihdk dk}cpkho+
Mhohi qhpdhud ic}gmc/ ^pg}glgo ^EP }djh ohkjlhb mdjskhlhk sk}sl ~ci`ck}slhk ~pg}cdk
ebdicpde/ bhkth uh}s ohkjlhb logkdkj md`s}sblhk sk}sl `cp}slhp mhpd mgihdk dk}cpkho "Jphkmgpd
c} ho 844:$+
Ic}gmc dkd mh~h} mdjskhlhk sk}sl ickjjhk}dlhk ush}s mgihdk }cp}ck}s mhohi ~pg}cdk H gocb
mgihdk hkhogj mhohi ~pg}cdk @+ U}smd }ck}hkj bd`pdmh mgihdk)`cp}slhp mh~h} mdbhudolhk +
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 3>
-
7/23/2019 Protein Engineering _ Siti Julaiha
51/98
IC]GMC [email protected]
Lhmhkj)lhmhkj usod} mdlc}hbsd is}hud uc~cp}d h~h thkj mh~h} ickjbhudolhk udah}
}cp}ck}s ~hmh ush}s ~pg}cdk
Iduhokth/ sk}sl ickdkjlh}lhk }cpigu}h`dod}hu ush}s ~pg}cdk ihudb usod} sk}sl
mdohlslhk bhkth mckjhk icohlslhk ~pcmdlud ~hmh u}psl}sp 5 mdickud "5)M$ sk}sl
ickck}slhk huhi hidkg ihkh thkj ickck}slhk udah} dkd
Mckjhk ic}gmh dkd mh~h} mdohlslhk is}hud ~hmh `cp`hjhd }ci~h} sk}sl
lcismdhk mdohlslhk dmck}dadlhud ~pg}cdk is}hk thkj ihkh thkj mh~h}
ickjbhudolhk askjud thkj mddkjdklhk
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 38
-
7/23/2019 Protein Engineering _ Siti Julaiha
52/98
Ickjjskhlhk ~ckmclh}hk dk qd}pg hehl / ~pcmdlud }cpbhmh~ aphjick ~pg}cdk thkj bhpsu
ickth}s mhk ~gudud epguugqcp thkj bhpsu md}ci~h}lhk }dmhl md~cposlhk+
Ic}gmc dkd icidodld `c`cph~h }clkdl ~ckmclh}hk thd}s 6
) ^ckmclh}hk ebdicphjckcudu hehl hmhohb ~cktdud~hk hehl+ Uc~cp}d khihkth/ ic}gmc
dkd icod`h}lhk ~cktdud~hk sps}hk MKH lc oglhud hehl jck }hpjc} "Bcaapgk c} ho+ 84:2< Oselgz c}
ho+ 842:< Bhooc} c} ho+ 844:< Ihkgdo mhk @hdoct 844:$+
) Dkqcu}djhud bd pdmh thkj mdbhudolhk gocb ~cktdud~hk hehl mh~h} mdjskhlhk sk}sl
ickjdmck}dadlhud ud}su)ud}su thkj thkj ickcpdih dkucpud askjudgkho mhpd uc`shb mgihdk ~pg}cdk
ickfhmd ~cphkehb'uehaagom " @c}}gk c} ho+844:$+
) ^ckmclh}hk ohdk/ ~cpis}hud icodkjlhp ~pg}cdk/ ickjbhudolhk pcoglhud K)mhk E)
]cpidkd "Jpha mhk Uebhebihk 8441$+ Ic}gmc dkd mh~h} mdjskhlhk sk}sl icihbhid fhosp
cqgosud mhk sk}sl ickjdmck}dadlhud ud}su ~cpiduda ucehph udu}cih}du sk}sl ~cpis}hud icodkjlhp/
dkagpihud ickjskjlh~lhk }ck}hkj igmsohpd}hu uehaagopm "@hdpm+ C} hoo 8444< Bckkcelc c}
ho+ 8444$+
) ^ckmclh}hk Pclgi dkhud bgigogj mhk
) ^ckmclh}hk Pclgi`dkhud kgkbgigogj
^ckfcohuhk ~ckmclh}hk ~ckmclh}hk dkd md~h~hplhk uc`hjhd `cpdls} 6
IC]GMC CKMCLH]HK EBDICPHJCKCUDU HEHL
Ucehph }cgpd/ }clkdl dkd uc`cpkhpkth mh~h} md`hjd ickfhmd ~ckmclh}hk kgklgi dkhud mhk
lgi`dkhud mckjhk lcsk}skjhk",$ mhk lcpsjdhk")$ thkj md~h~hplhk `cpdls} 6
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 30
-
7/23/2019 Protein Engineering _ Siti Julaiha
53/98
^hmh ic}gmc dk/d ^pg}cdk Bd`pdm thkj ickjhkmskj ucjick "mgihdku ' us`mgihdku$ mhpd ~hodkj
ucmdld} msh kh}spho thkj `cp`cmh h}hs ~pg}cdk mhpd gphkj }sh ocohld "Hpiu}pgkj 844>$+ Mgihdku
mhk us`mgihdku mdmcadkdudlhk }cpbhmh~ u}psl}spho ig}da sk}sl qhpdhud slsphk mhk lclgi~oclhk/
}cpihusl skd} skd} lcedo thkj ph}h ph}h icidodld 8> 5> huhi hidkg/ icod~h} ickfhmd skd}
askjudgkho / mhk mgihdku `cuhp mhpd `c`cph~h ph}su huhi hidkg thkj icidodld hl}dqd}hu ckvdi
"Gu}cpicdcp hkm @cklgqde 0>>8$+
@c`cph~h }d~c bt`pdmu6
) Udkjoc epguugqcp bt`pdmu }cpmdpd mhpd `hjdhk K)}cpidkho uh}s ~pg}cdk mhk `hjdhk E)
}cpidkho ~pg}cdk ohdkkth+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 35
-
7/23/2019 Protein Engineering _ Siti Julaiha
54/98
) Iso}d~oc epguugqcp bt`pdmu/ uh}s h}hs oc`db dk}cpkho u}pc}ebcu mhpd ususkhk huhi
hidkg md}ci~h}lhk ~hmh ucjick thkj `cpbs`skjhk mhpd ckvdi ohdk+
) Asudgk bt`pdmu/ mgihdk askjudgkho mhpd ~pg}cdk thkj }cpbs`skj mckjhk mgihdk mhpd
~pg}cdk ohdkkth/ icked~}hlhk ~pgmsl jh`skjhk'asuc thkj oc`db `cuhp mhpd ~hpck}kth+
Bt`pdm mh~h} icktc`h`lhk is}hud/ ~ckjbdohkjhk mhk ~ckhi`hbhk huhi hidkg+
Ph}dgkho mcudjk bt`pdmu ici`s}sblhk dmck}dadlhud ~pcodidkhpd mhpd igmso askjudgkho "Bg~akcp c}
ho+ 8442< Uebkcdmcp c} ho+ 8442$ mckjhk dku~clud }cpbhmh~ u}psl}sp 5 Mdickud ~hmh uc}dh~ ckvdi
huhokth+
]dmhl hmhkth dkagpihud u}psl}sp/ mh~h} mdjhk}dlhk mckjhk dkagpihud hodjkick} ususkhk huhi
hidkg sk}sl ickjdmck}dadlhud ucjick ~g}ckudho thkj ~ck}dkj+
Ususkhk MKH mddku~clud ~hmh oglhud ~cpislhhk ogeh}c crgkdk}pgk thkj ickmcadkdudlhk `h}hu
u}psl}sp mhk askjud skd}+
Ic}gmc Hmqhkecu bt`pdm icked~}hlhk ebdicpde od`phpdcu ~hmh }d~c phkmgi/ ickjbdohkjlhk
lc~cposhk h}hu dkagpihud u}psl}sp h}hs ususkhk hodjkick} "Gu}cpicdcp c} ho+ 8444`< U}cqckugk
hkm @cklgqde 0>>0$+
Phkmgi ic}bgmu iciskjldklhk sk}sl icihi~hplhk lgk}pd`sud ucjick ~pg}cdk }cpbhmh~
askjudkth }hk~h ~pcegkecdqcm dhu+
Ic}gmc ici~skthd lcsk}skjhk }cpbhmh~ }d}dl jh`skjhk hk}hph mgihdk u}psl}sp mhk askjud h}hs
us`skd} mh~h} mdoglhudlhk mckjhk ucushd+
Dku~clud bt`pdmu mh~h} icked~}hlhk ~ckjcp}dhk thkj `cpbhpjh }cpbhmh~ cosudqc mhk pcudmsc
~ck}dkjpcudmsc egk}he}u mhk ogkj)mdu}hkec pcudmsc kc}zgplu thkj uhkjh} lpsudho ~hmh hl}dad}hu
~pg}cdk "Hjhpzho c} ho+ 0>>0< Phfhjg~hohk c} ho+ 0>>0< @cklgqde hkm Bhiicu)Uebdaacp 0>>5$+
IC]GMC [email protected] BGIGOGJ
Pclgi`dkhud bgigogj mhpd aphjick jck ickhkmhlhk pchuuci`ot aphjick MKH mhpd jck
gphkj}sh mckjhk ickjjh`skjlhk MKH mhpd `c`cph~h gphkj }sh ickfhmd ~pgmsl hlbdp jck+
Iclhkduic pchuuci ot mdmhuhplhk ~hmh sps}hk MKH bgigogjd mhk jck mckjhk bgigogjd
pckmhb "lsphkj mhpd :>#$ }dmhl mh~h} mdlgi`dkhudlhk "Udc`cp c} ho+ 0>>8$+ Hld`h}kth/ hmh `dhu
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 3=
-
7/23/2019 Protein Engineering _ Siti Julaiha
55/98
dkbcpck mhohi pclgi`dkhud bgigogj od`phpdcu6 ~gudud epguugqcp fh}sb ~hmh mhcphb }dkjjd/ `slhk
pckmhb/ lcuhihhk MKH +
Iculd~sk mcidldhk/ ic}gmc dkd mh~h} uhkjh} `cpjskh mhohi mgihdk uzh~~dkj/ lhpckh
ucpdkjlhod jck gphkj}sh `cp`hjd bgigogjd }dkjlh} }dkjjd+ Ucohkfs}kth/ pclgi`dkhud bgigogj
mh~h} mdjskhlhk sk}sl ickjjh`skjlhk aphjick mhpd oc`db mhpd msh jck dkmsl/ iciskjldklhkickfcohfhbd sps}hk mhpd ucospsb lcoshpjh ckvdi mhohi uh}s ~cpeg`hhk+
@cpdls} hmhohb Lkglgs}u Jck thkj mdbhudolhk gocb pclgi`dkhud bgigogj 6
Ic}gmh sk}sl iciduhblhk ~pg}cdk mckjhk udah} thkj mddkjdklhk mhk ickjdmck}dadlhud is}hud
~cktc`h` ~cps`hbhk6
8+ ^ckcosusphk "Ulpdkdkj$6 ucish ~su}hlh thkj hmh md~cpdluh ucehph hehl sk}sl ickehpd
~pg}cdk mjk udah} thkj mddkjdklhk
0+ Ic}gmh ucoclud6 ^su}hlh mducoclud ucmcidldhk ps~h ucbdkjjh bhkth thkj icidodld
~cp`hdlhk udah} thkj md}cosuspd
^pg}glgo md}cph~lhk ~hmh ic}gmh lgi`dkh}gpdho6
8+ Ic}gmh ~ci`sh}hk ~su}hlh lgi`dkh}gpdho mckjhk lcphjhihk thkj }dkjjd ucbdkjjh
ickdkjlh}lhk lciskjldkhk md~cpgocb ~pg}cdk mckjhk udah} thkj mddkjdklhk
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 33
-
7/23/2019 Protein Engineering _ Siti Julaiha
56/98
0+ ^ckck}shk ic}gmh thkj mdjskhlhk sk}sl iciduhblhk ~pg}cdk mckjhk udah} thkj
mddkjdklhk mhk ickjdmck}dadlhud is}hud ~cktc`h` ~cps`hbhk
IC]GMC [email protected] BGIGOGJ MKH UBSAAODKJ
^pg}glgo mhuhp sk}sl pclgi`dkhud bgigogj dk qd}pg bgigogj jck gphkj}sh mduc`s} uc`hjhd MKH
Ubsaaodkj "Zdoodhi ^E U}ciicp/ 844=/ 8442$+
MKH UBSAAODKJ hmhohb ]clkdl cqgosud mckjhk lhphl}cpdudl `cp`cmh thkj ickdps ~pgucu
mcuhdk h}hs pclgi`dkhud hohid ~hmh pc~pgmslud uclusho mhk ici~cpec~h} ~pgucu
~cphkehkjhk icohosd ucoclud ohkjuskj ~hmh dk qd}pg thd}s ickehi~splhk mhk icktcushdlhk
~pg}cdk "ih}eb$ sk}sl ickjbhudolhk qhpdhk oc`db/ mckjhk ici~cp}di`hkjlhk 6
) ]clkdl dlh}hk'Agomdkj ^pg}cdk
) ]cpigu}h`dod}hu
) Hl}dqd}hu Ckvdi
H~odlhud }clkdl dkd mdohlslhk ~hmh Pclgi`dkhud bgigogj dkqd}pg mhpd `c`cph~h ucoclud ^ggo Jck
icohosd Aphjick}hud hehl h}hs Phkmgi mhk ~cipgucuhk lci`hod icohosd }clkdl ^EP+
Uclcogi~gl bgigogj jck ucehph hehl md`cohb ickfhmd aphjick lcedo mckjhk ~ckjbhkesphk
gocb Ckvdi pcu}pdlud MKhucD+
Aphjick ucohkfs}kth md~huhkj lci`hod gocb ^EP md ihkh icpclh `cpaskjud uc`hjhd }ci~oh}cmhk ~pdicp+ Aphjick icktcohphulhk mhk epguu)~pdic uh}s uhih ohdk sk}sl pc~odlhud
ici`cpdlhk sk}hd MKH bd`pdmh mckjhk `c`cph~h lgi~gkck mhpd jck dkmsl thkj huod+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 31
-
7/23/2019 Protein Engineering _ Siti Julaiha
57/98
MKH ubsaaodkj icidi~dk ~cplci`hkjhk }clkdl }cplhd} thkj ickhzhplhk lcskjjsohk mhohi
ud}shud }cp}ck}s+ Iduhokth/ mhohi igmdadlhud ucmcpbhkh mhpd ~pg}glgo huod/ mdmhuhplhk ~hmh
ckvdi pcu}pdlud ubsaaodkj/ Ldlsebd c} ho+ "8444$ mdjskhlhk ehi~sphk pcu}pdlud ckmgkslochuc
uc`hjhd jhk}d MKhucD sk}sl icig}gkj jck dkmsl+
Bho bho thkj md~cposlhk sk}sl icohlslhk pclgi`dkhud Bgigogj MKH Ubsaaodkj dkd hmhohb 6
) ^ckcph~hk pclgi`dkhud bgigogj ~hmh ususkhk ususkhk thkj oc`db mhpd 8 L`
) Lc`cphmhhk mhcphb Bgigogjgsu iciduhblhk mhcphb mhcphb thkj `cp`cmh "~cp`cmhhk
zdohthb$+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 3:
-
7/23/2019 Protein Engineering _ Siti Julaiha
58/98
) U}psl}sp Uehaagpm)odlc ^pg}cdk thkj iciskjldklhk ~pgucu thkj ucushd sk}sl Ubsaaodkj '
^ckehi~sphk+
^ckjci`hkjhk Ic}gmc Pclgi`dkhud Bgigogj mckjhk MKH Ubsaaodkj dkd ickjhohid `hkthl
~cplci`hkjhk mhohi h~odlhudkth md `dmhkjk Pclhthuh ^pg}cdk/ mdhk}hphkth 6
8+ Ahidot Ubsaadkj
Thd}s jck `cphuho mhpd ahidot thkj uhih/ icps~hlhk bgigogj thkj }dkjjd+
Egk}gb ubsaaodkj dkd mdohlslhk ~hmh pclhthuh jck Ebc~bhogju~gpdkhuc ickfhmd
Hk}d`dg}dl Ohe}hi "Hl}dqd}hu Igrhohl}hihuc$ mdihkh Logk thkj }cp`hdl ickjhkmskj }d}dl
is}hud mcoh~hk ucjick mhpd 5 ~cp = jck mhk 55 huhi hidkg + Ickdkjlh}lhk
lc}hbhkhk 0:>)3=> dlh}hk'agom
Egk}gb mhpd ahidot ubsaaodkj mckjhk cadudckud }dkjjd `cpmhuhplhk iso}d u}c~ ^EP mhk
Pclgi`dkhud MKH dk qdqg mdohlslhk ~hmh Tchu} ' Fcphid / mckjhk ickjhooduh askjud mhk
ickjsi~solhk u}h}du}de mh}h mhpd lgi dkhud ^su}hlh hk}hph Bsihk Et}gebpgic
^=3>8H8 mhk 8H0/ thkj mdlci`hkjlhk gocb Mckdu ^gi~gk/ Jdoocu ]pshk mhk Qhocpdc
H`cehuudc/ 0>>>+ H~odlhud dkd mdlckho mckjhk uc`s}hk Ubsaaodkj Lgi dkhu Od`phpdcu
Ckbhkecm gocb Pclgi`dkhud ~hmh Tchu} h}hs mdudkjlh} EOCPT+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 32
-
7/23/2019 Protein Engineering _ Siti Julaiha
59/98
0+ Ckvdi Pcu}pdlud Ubsaaodkj Ebdihpchu
Md`hkmdkjlhk mckjhk ~pg}glgo MKH ubsaaodkj huodkth/ Ckvdi pcu}pdlud ubsaaodkj icidodld
lcsk}skjhk ickjbhudolhk apclsckud ebdicphu thkj oc`db }dkjjd lhpckh mdbdkmhpdkth
~ckjjskhhk MKhucD+
Bdmpgodudu MKhucD ~hmh MKH `cpsk}hd jhkmh oc db icidodb oglhud thkj `cpmclh}hk
mckjhk kslocg}dmh ~dpdidmdk mhk hld`h}kth `duh icisohd sps}hk `dhu lc pclgi`dkhud
}cpuc`s}+
Pcu}pde}dgk)ckvdi `cp`hudu ubsaaodkj/ici~skthd lcocihbhk `hbzh ud}su epguugqcp `dhu
`cp}c~h}hk mckjhk ~ci`h}huhk ud}su)ud}su thkj usmhb hmh+
Egk}gb ubsaaodkj dkd mhohi pclhthuh ~pg}cdk iduhokth ~hmh Bs)DAKu thkj mdlgmclhk
gocb ahidot mhpd oc`db 0> ms~odlh} jck kgk hoocode ici`hjd 23)42# sps}hk dmck}d}hu ~hmh
}dkjlh} huhi hidkg+
^pg}cdk dkd icidodld hl}dad}hu hk}dqdpho mhk hk}d~pgodacph}dqc thkj lsh} thkj icidodld s}dod}hu
lodkdu uc`hjhd hk}dlhklcp mhk }cph~d hk}dqdpsu+ Iculd~sk s}dod}hu DAKu ebdicpde `cphuho
mhpd lcoshpjh jck dkd }cohb mdhlsd/ bhkth uc`hjdhk lcedo mhpd lciskjldkhk 8>[01
ebdicphu }cohb mdclu~ogphud/ `hdl mhohi cqgosud ihksudh hohid h}hs mckjhk ic}gmc
`dgogjd igoclsocp igmcpk/ mhk bhkth uh}s DAK us`}t~c)hohi/ Bs) DAK)0/ }cohb
mdjskhlhk mhohi u}smd lodkdu+ DAK thkj ~hodkj hl}da mdpclhthuh hmhohb DAK hoahegk)8/
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 34
-
7/23/2019 Protein Engineering _ Siti Julaiha
60/98
hmhohb uc`shb lgkuckusu mhpd 85 zdom }t~c)jck Bs)DAK thkj uhh} dkd mdjskhlhk mhohi
}cph~d bc~h}d}du E+
5+ U}hjjcpcm Cr}ckudgk pgecuu "U}C^$
hmhohb qhpdhud ohdk mhpd MKH ubsaaodkj "Vbhg c} ho 8442>>$+
Ic}gmc dkd mdjskhlhk sk}sl icked~}hlhk ~pg}cdk ~pg}cdk bt`pdm+
Mhohi ~pgucmsp dkd/ }cpidkho ~pdicp mdjskhlhk sk}sl ickdps MKH }hpjc} ickjjskhlhk
^EPu mckjhk clu}ckud zhl}s thkj uhkjh} udkjlh} uclhod sk}sl ickjbhudolhk sk}hd
~ckmcl MKH thkj mdpc~odlhud+
Phk}hd }si`sb MKH `cp}dkmhl uc`hjhd ~pdicp mhohi udlosu `cp}sps})}sps} mhk ~cps`hbhk
}ci~oh}c mdpc~odlhud `c`cph~h lhod/ mckjhk mcidldhk ickjsi~solhk Lgi~gkck mhpd jck
dkmsl thkj `cp`cmh+
^hmh ~pgucu dkd md`s}sblhk ususkhk MKH mckjhk bgigogj thkj }dkjjd+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 1>
-
7/23/2019 Protein Engineering _ Siti Julaiha
61/98
Egk}gb h~odlhud ~hmh ubsaaodkj dkd ~ckdkjlh}hk bhoa)odac sk}sl us`u}dock+
Jck bd`pdm mdlgku}psludlhk mhpd msh jck bgigogj+ Eogkdkj mhpd jck bd`pdm ickjbhudolhk
~pgmslud ~pg}cdk bd pdm+ Bhkth uh}s phk}hd mhpd uc}dh~ jck md}skfsllhk uc}cohb ohkjlhb
mckh}sphud hzho+
=+ Pclgi dkhud ~pdidkj hehl "PHEBD]]$
Qhpdhud ohdk mhpd MKH ubsaaodkj sk}sl icked~}hlhk bt`pdm ~pg}cdk hmhohb 6
) Pclgi`dkhud ~pdidkj hehl "Ubhg c} ho 8442$+
Thd}s }clkdl pchuuci ot '~ckjsi~sohk MKH mckjhk udk}cuh dk}cpps~}dkj'~cktcohhk thkjmdclu~cpdick}lhk gocb Ubgp} 844:+
MKH Ubsaaodkj mhk ic}gmc thkj }cplhd} }cohb mdh~odlhud ~phl}du md `dmhkj pclhthuh
~pg}cdk dkmsu}pd/ }c}h~d bhudokth ucpdkjlhod usod} sk}sl icphudgkhoduhud lhpckh ebdicphu
thkj mdbhudolhk icidodld hkthl ~hpck}u mhk c`cph~h ~gudud epguugqcp+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 18
-
7/23/2019 Protein Engineering _ Siti Julaiha
62/98
Bho dkd }dmhl udjkdadlhk `doh }sfshkkth hmhohb ~pgmsl hlbdp/ thd}s ckvdi mdpclhthuh
mckjhk udah} thkj mddkjdklhk+
) Pclgi`dkhud Ebdicphjckcudu hehl ~hmh }ci~oh}c }phkudck h}hs ucick}hph
"PHEBD]]$ thkj mdclu~cpdicklhk gocb EgEg c} ho+ 0>>8+
3+ Dkepcick}ho ]pskeh}dgk agp ]bc Epch}dgk Bt`pdm Ckvticu "D]EBT$
Hmhohb ic}gmc ~ckmclh}hk lgi`dkhud sk}sl icked~}hlhk bt`pdm mhpd msh ~pg}cdk thkj
`cpbs`skjhk+ Msh ~pg}cdk thkj `cpbs`skjhk mdcklgmclhk gocb jck 8 mhk jck 0 + Bhudo
hlbdp kth icps~hlhk jck bd`pdm thkj }cpmdpd mhpd sfskj 3 jck 8 mhk sfskj 5 jck 0+
]clkgogjd ~cig}gkjhk }hi`hbhk ickjdobhid ~ckcishk u}ph}cjd ebdicphjckcudu thkj
oc`db ihfs mhk oc`db ucushd sk}sl icihbhid bs`skjhk u}psl}sp mhk askjud ~pg}cdk+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 10
-
7/23/2019 Protein Engineering _ Siti Julaiha
63/98
U}ph}cjd ~cp}hih mhpd ~cig}gkjhk dkd/ hmhohb ~cig}gkjhk dkepcick}ho sk}sl
~ci`sh}hk ckvdi bd`pdmh "D]EBT)Dkepcick}ho }pskeh}dgk agp }bc epch}dgk Bt`pdm
Ckvticu "D]EBT$ thkj mdlci`hkjlhk gocb J/ ]pshk c} ho+/ 0>>>/ ickjbhudolhk
~cp~su}hlhhk ~pg}cdk bd`pdmh udkjoc epguugqcp hk}hph msh jck dkmsl+
H~odlhud bhudo D]EBT }cpmh~h} ~hmh ~cp~su}hlhhk jck mckjhk ~cig}gkjhk }hi`hbhk~hmh ~gudud epguugqcp thkj mdmdu}pd`sudlhk ucehph hehl+
^cp~su}hlhhk dkd `cpdud bd pdmh mckjhk udud~hk mhk ~ckjbh~suhk MKH/ mhk ~pgmsl)
~pgmsl mhpd cp`hjhd slsphk thkj md~pgmslud+
]hi`hbhk ~cig}gkjhk "hppgz ~cp}hih$ mhk ~ckhi`hbhk ~cig}gkjhk ckvdi bd`pdmh
"D]EBT$+ Mhohi D]EBT/ ~cp~su}hlhhk ~cig}gkjhk dklpcick}ho mdbhudolhk gocb
~ckjbhkesphk MKH gocb crgkseochuc DDD+
Aphjick thkj mdbhudolhk lcismdhk mdodjhudlhk `cpuhih)uhih sk}sl ickjbhudolhk
~cp~su}hlhhk hlbdp+
]bdg)D]EBT/ icps~hlhk qhpdhud ic}gmc D]EBT/ mckjhk ickjjskhlhk Hkhogj
kslocg}dmh }pdaguah} thkj mdlci`hkjlhk gocb Os}v/ c} ho+0>>8+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 15
-
7/23/2019 Protein Engineering _ Siti Julaiha
64/98
Mhohi uc`shb ^EP/ mK]^ )~bgu~bg}bdgh}c mdjh`skjlhk ucehph hehl mhk ~hmh
apclsckud pckmhb lc mhohi zdohthb thkj md}hpjc}lhk sk}sl ~cig}gkjhk MKH+
Dk}cpkseocg}dmc ~bgu~bg}bdgh}c thkj mdbhudolhk ickjbs`skjlhk lc}hbhkhk }cpbhmh~
bdmpgodudu crgkseochuc 5*)3 */ ici`cpdlhk lc}hbhkhk MKH }hpjc} }cpbhmh~ mcjphmhud
mhohi ~ci`sh}hk CrgD `cpdls}kth+
]bdg)D]EBT }cohb `cpbhudo mdjskhlhk sk}sl ickjjh`skjlhk lci`hod jck mckjhk sps}hk
dmck}d}hu mckjhk bhkth 51# sk}sl ickjbhudolhk ebdicphu hl}da "Uhpha c} ho 0>>1$
Lc}cp`h}huhk s}hih D]EBT mhk ]bdg)D]EBT hmhohb `hbzh epguugqcp bd`pdmh }skjjho
bhkth mh~h} mdbhudolhk mhpd msh gphkj }sh / ici`h}hud ~g}ckud lchkclhphjhihk
sps}hk+
1+ ^ckjjh`skjhk' Asudgk D]EBT mhk MKH Ubsaaodkj ucehph `cpsps}hk
"UEPH]EBT$
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 1=
-
7/23/2019 Protein Engineering _ Siti Julaiha
65/98
Ic}gmc pclgi`dkhud bgigogj `cpbhudo mdjskhlhk mckjhk ickjlgi`dkhudlhkkth ~hmh
ic}gmc ohdk sk}sl ickck}slhk ahl}gp)ahl}gp ~ckck}s u~cudadud}hu us`u}ph} mhk hl}dad}hu
ckvdi ~hmh `cp`hjhd qhpdhud ahidot ~pg}cdk+ "Jpduzgom c} ho+ 0>>3< ^hpl c} ho+ 0>>1$+
Ic}gmc dkd mdjskhlhk sk}sl ickehpd bs`skjhk u}psl}sp)askjud mhk iclhkduic cqgosud
~pg}cdk "Jpduzgom c} ho 0>>3>1$+
Lc}dlh mdlgi`dkhudlhk mckjhk ulpdkdkj jckc}dl h}hs }clkdl ~dodbhk/ mh~h} mdjskhlhk
sk}sl ici~cpgocb zhzhuhk hps lc mhohi ckvdi+
Sk}sl ici`sh} ~cp~su}hlhhk `c`cph~h epguugqcp/ @cklgqde mhk pclhk lcpfh
ickjjh`skjlhk D]EBT mhk MKH ubsaaodkj ucehph `cpsps}hk mhohi ic}gmc dkd mduc`s}
UEPH]EBT "Os}v c} ho 0>>8`$+
:+ Bgigogjd Sps}hk Dkmc~ckmck pg}cdk "UBD^PCE$
Udc`cp mhk pclhk lcpfh ickjci`hkjlhk }clkdl thkj `cp`cmh sk}sl icked~}hlhk
epguugqcp uh}s ~cp~su}hlhhk ebdicpde hk}hph msh jck ~hpck}ho }hk~h `dhu sk}sl ~gudud
epguugqcp "Udc`cp c} ho+ 0>>8$+
Ic}gmc pclgi dkhud bgigogjd sps}hk dkmc~ckmck ~pg}cdk h}hs Ucxsckec
Bgigogjt Dkmc~ckmck} ^pg}cdk Pcegi dkh}dgk"UBD^PCE$/ mdisohd mckjhk ~pgmslud
ush}s mdicp jck thkj lcismdhk }cp~cehb)~cehb gocb MKhucD+
Aphjick} slsphk gphkj}sh md~sodblhk mhk md`hodllhk gocb odjhud icodkjlhp mhk pcu}pde}dgk
ickjbhkesplhk sk}sl ickjbhudolhk uc`shb ~cp~su}hlhhk ckvdi bd pdmh mckjhk
mdu}pd`sud thkj icph}h mhpd ~gudud epguugqcp+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 13
-
7/23/2019 Protein Engineering _ Siti Julaiha
66/98
2+ CRGK Ubsaaodkj
Dk qd}pg crgk ubsaaodkj icps~hlhk crgk mgihdk ckegmdkj h}hs uc} mhpd crgk ~hmh }hpjc}
jck mhk bgigogj mhpd thkj ohdkkth+ Jck thkj `cpbs`skjhk mdhi~odadlhud
ickjjskhlhk ebdicpde godjgkslocg}dmc ~hmh ~pdicp mhk }ci~oh}c ~hmh pchlud uc~cp}d
^EP thkj icktsusk lci`hod Jck mckjhk ~hkfhkj ~cksb+ U}c~ }cphlbdp icod`h}lhk~ckthpdkjhk ^su}hlh Crgk ubsaaodkj sk}sl askjud h}hs ~pg~cp}t thkj mddkjdklhk+
]clkdl dkd mdlci`hkjlhk gocb Fggu} H Lgolihk mhk Zdooci ^+E+ U}ciicp/ 0>>8+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 11
-
7/23/2019 Protein Engineering _ Siti Julaiha
67/98
IC]GMC [email protected] KGK BGIGOGJ
Uc`hjhdihkh md`hbhu/ Lc}cp`h}huhk s}hih ic}gmc pclgi dkhud bgigogj hmhohb `hbzh
bhkth jck mckjhk bgigogjd }dkjjd thkj mh~h} mdlgi`dkhudlhk+
Ucpdkjlhod lcogi~gl ~pg}cdk ici`hjd u}psl}sp }djh)mdickud thkj uhih/ }c}h~d sps}hk MKH
dmck}d}hukth pckmhb+
Pclgi dkhud uc~cp}d lcogi~gl ~pg}cdk mckjhk ic}gmc pclgi dkhud bgigogj hlhk
ickjbhudolhk uc`shb ~cp~su}hlhhk md ihkh ~gudud epguugqcp hlhk `cp`hpdkj ucehph clulosuda
~hmh mhcphb lcedo ususkhk MKH mckjhk bgigogjd }dkjjd+
Uc`hodlkth/ ic}gmc pclgi`dkhud kgk bgigogj }dmhl `cpjhk}skj ~hmh sps}hk bgigogjd jck
gphkj }sh lhpckh }dmhl icohlslhk bd`pdmduhud aphjick bgigogj / icohdklhk ici`sh} ohkjlhb
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 1:
-
7/23/2019 Protein Engineering _ Siti Julaiha
68/98
odjhud `cpsfskj }si~so "`osk})ckmcm$ thkj mdjskhlhk sk}sl ici`hzh jck aphjick `cpuhih+
Hld`h}kth/ }dmhl hmh `dhu }cpbhmh~ lgi~gudud aphjick jck md}cisd+
Mckjhk ic}gmc pclgi`dkhud kgkbgigogj/ iciskjldklhk sk}sl ici`sh} ~cp~su}hlhhk
ebdicphu mhpd msh jck thkj uc~cksbkth }dmhl `cpbs`skjhk+
Uc`hjdhk `cuhp ic}gmc pclgi`dkhud kgkbgigogj mdmhuhplhk ~hmh ~cig}gkjhk dklpcick}ho/
mhohi `ck}sl thkj ~hodkj ucmcpbhkh ickskfsllhk ~cked~}hhk ~cp~su}hlhhk ~pg}cdk thkj
icidodld uh}s h}hs oc`db huhi hidkg / mdbh~su `hdl mhpd E)h}hs K)}cpidksu +
"Gu}cpicdcp c} ho 8444h$+ ^cig}gkjhk ]hi`hbhk mhk ic}gmc }cplhd} }cpjhk}skj ~hmh
crgkseochuc DDD "CrgDDD$ ~pg}cdk mhk udah})udah}kth+
CrgDDD ickjlh}hodudu ~ckecpkhhk aphjick jck mhpd 5 *lc sfskj 3* ~hmh mdlgk}pgo/ ucphjhi/ mhk
udklpgk }dkjlh} "Zs c} ho 84:1$+ Hodxsg}u Lcedo ehi~sphk pchlud mdbh~su ucohih ohkjlhb
~ckecpkhhk sk}sl ici`sh} ~cp~su}hlhhk jck mckjhk kgigp thkj `cp`cmh mhpd ~huhkjhk
`huh MKH thkj mdbh~su+
H^ODLHUD IC]GMC [email protected]
g Phkehkjhk pclgi`dkhk Bcigjog dk ihksudh mdjskhlhk sk}sl ickck}slhk ~ckjjhk}d
mhphb+ Mdohlslhk gocb Ogglcp M/ H``g}})@pgzk M/ Egvhp} ^/ Mspacc U/ Bgaaihk U/ Ih}bczu HF/
Idoocp)Pgcbpdeb F/ Ubgcihlcp U/ ]pdi`oc U/ Acpid J/ c} ho Kh}spc 5316032)1> / "8440$
g Cacl ^ckhkhihk hoogu}cpdl skdl mhpd `shth lc~hmh bcigjog`dk ihksudh/ Lgidthih KB/
Idthvhld J/ ]hic F/ Khjhd L Kh}spc 5:560==)1 "8443$
g Icpclhthuh is}hk BJA'UA mckjhk dlh}hk thkj mdlsphkjd sk}sl bc~hpdk usoah} ~pg}cgjodlhk
mhk ickdkjlh}lhk hl}dad}hu dk qdqg+ Mdohlslhk gocb Bhp}ihkk J+/ ^pgu~cpg ]+/ @pdklihkk Q+/
Gvecodl G+/ Zdk}cp J+/ Bc~~oc F+/ @h}oct U+/ @ohm} A+/ Uhebu I+
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 12
-
7/23/2019 Protein Engineering _ Siti Julaiha
69/98
H^ODLHUD MCUHDK PHUDGKHO CQGOSUD THKJ MDHPHBLHK"MDPCE] CQGOS]DGK$
Ickdkjlh}lhk ckvdi ad}kcuu sk}sl h~odlhud thkj md}ck}slhk
Ckvtic hmh~}hud6
Ickdkjlh}lhk hl}dad}hu Us`u}ph} @hps
Us`u}ph}c u~cedaded}hu )) Iso}d~oc us`u}ph}cu haadkd}t
]bcpigu}h`dod}hu)) ico}dkj ~gdk} "]i$
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 14
-
7/23/2019 Protein Engineering _ Siti Julaiha
70/98
Hmh~}hud ]ci~cph}sp6 He}dqd}hu ~hmh usbs thkj cp`cmh
)) ^utebpg~bdode qu+ Icug~bdode qu+ ]bcpig~bdode
U}h`dod}hu h}hs hl}dad}hu ~hmh odklskjhk hp}dadedho'`sh}hk
)) Ugoqck} qu+ hxscgsu ugos}dgku
)) Ic}ho dgku "~pg}chuc$
Hk}d`dg}de pcudu}hkud
Ldpd 6 Od`phpd jckcphud is}hk hehl ckvdi ]CI)8 ickjjskhlhk ^ mhk 2 )grgJ is}hjckcudu
Lhkhk6 H)]CI 8 is}hk "5M+3$ thkj mdbhudolhk gocb phkmgi is}hjckcudu mckjhk 0+3>> lhod
od~h} Hl}dqd}hu bdmpgodudu thkj oc`db }dkjjd }cpbhmh~ hk}d`dg}de ucahogu~gpdk "mhpd Vheegog mhk
Jbcphpmd/ F igo @dgo 8444$+
Egk}gb
U}smd cqgosud thkj mdhphblhk ~hmh hmh~}hud }ci~hphp}sp Ckvdi ^udlpgadodl
"Lck}hph Idthvhld/ ^h}pdel Zdk}pgmc/ Pgzhk Jphtodkj/ Mgkk Ps`dkjb/ Aphkecu Hpkgom/ 0>>>$
) Ususkhk Huhi hidkg Hodjkick} ~hmh Us`}dodudk U=8"Mhqhdo/ c}+ho 8440$/ U54 "Khpdkr/
8440$/ UUDD "Zh}d/844:$/ @^K "Zcoou/ 8425$/ C "U}hbo Acpphpd /2=$/Ehpou`cpj "Fheg`u/23$
Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc :>
-
7/23/2019 Protein Engineering _ Siti Julaiha
71/98
mhk ]cpid}huc "Icogsk/ 23$+ Hodjkick iso}d~oc mdohlslhk mckjhk EOSU]HL Z
"]bgi~ugk/4=$+ Pcudms md}cislhk apclsck ;3># + Is}hud ~hmh 5)0J:+
) Odkchjc us`u}dodudk qhpdhk U=8+ Qhpdhk jckcphud ~cp}hih thkj mdmh~h} uc`cosikth+
"Idthvhld!Hpkgom/ 8444$+ ]sfsb is}hud mdmh~h} mhpd Jckcphud ~cp}hih mdpclgi dkhudlhk
gocb U}CG mhk qhpdhk }cp`hdl/ 0)1E3 mdmh~h} ~hmh jckcphud ^su}hlh lcmsh+ Md~hpck}lhk'mdbs`skjlhk mckjhk jck gphkj}sh sk}sl icked~}hlhk jckcphud ~su}hlh lc}djh gocb ^EP
Cppgp ~pgkc+ Bhkth kgkudkgkdi is}hud md}skfsllhk+
) Us }dodudk mhpd u}psl}sp thkj mdlc}hbsd+ ]sfsb is}hud }cpigu}h`do mddmck}dadlhud ~hmhm
qhpdhk 5)0J:+ Lhoudsi Eh bd}hi ickdkmdlh}c ud}c dlh}hk Eh0, thkj lc}h} mhk dkmdlh}c Eh
h`s h`s ud}c ickjdlh} Eh0, thkj ocihb/ thkj mdoglhudlhk md cohlhkj ogg~ thkj
ickjhkmskj Ucp8:3 Igmco mded~}hlhk ickjjskhlhk IGOUEPD^] "Lphsodu/ 8448$+
Mc~ckmhud ]ci~cph}sp hl}dad}hu u~cudadl us`}dodudk U=8/ 5)0J:/ mhk UU88+
) @hjhdihkh Ckvdi `cphmh~}hud mdodbh} mhpd u}smd cqgosud thkj mdhphblhk 6 Mdqcpjck}
cqgos}dgk ochmu sk}sl Hmh~}hud ~hmh odkjlskjhk thkj `cp`cmh / Uc`shb }phmc gaa hk}hph
hl}dqd}hu lh}hod}dl ~hmh usbs mhk }bcpigu}h`dodo} pckmhb+ "Idthvhld/ Zdk}pgmc$
) Pclhthuh U}h`dod}hu Ckvdi ]= odugvdc mckjhk dk}pgmslud ~hmh Dlh}hk U)0 "Ih}usisph$
) Pclhthuh U}h`dod}hu Ckvdi ]pdguc~bgu~h}c dugicphuc mhpd Tchu} mckjhk
ickjjhk}dlhk Huk "mchidkhud ~hmh }ci~cph}sp }dkjjd$ / Hbcpk c}+ho+
U}h`dod}h Ckvdi mdclu~pcudlhk uc`hjhd bhoa)odac/ h}hs ohfs dkhl}dqhud ckvdi ~hmh 8>>gE+ Bhoa
odac thkj oc`db ohih ickskfsllhk ckvdi thkj oc`db u}h`do+
) Pclhthuh Hl}dad}hu Ckvdi ]tpguto)}PKH utk}bc}huc mhpd @+ U}chpg}bcpig~bdosu mckjhk
ickjjhk}dlhk ]bp 38 "ickdkjlh}lhk haadkd}hu H]^$/ Zdoldkugk c}+ho+
) Pclhthuh ~ckjhpsb Eh' Ehoudsi sk}sl ickjhkhoduh dkmc~ckmckud Us`}dodudk+
Uh}sphud is}hjckcud / ickjbhudolhk : mhpd 8> mhcphb md}cislhk sk}sl ici`cpdlhk
~ckdkjlh}hk u}h`dod}hu+ Is}hk thkj mdbhudolhk 8>r oc`db u}h`do mhpd~hmh ckvdi huod mhohi
lc}dhmhhk