170228 Hendrix CV - Universiteit Hasselt Bioimaging Lab/1… · BOK protein multi-angled analysis...

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Jelle Hendrix | CV 1 Personalia Name Jelle Hendrix Day/Place of birth April 28, 1983, Hasselt Marital status married Children two UHasselt address Onderzoeksgroep Fysiologie Vakgroep Fysiologie-Biochemie-Immunologie Faculteit Geneeskunde en Levenswetenschappen Universiteit Hasselt Agoralaan Gebouw C – BIOMED, kantoor C034, B-3590 Diepenbeek tel: +32 11 269213 KU Leuven address Laboratory for Photochemistry and Spectroscopy Division of Molecular Imaging and Photonics Department of Chemistry Science, Engineering & Technology Group KU Leuven Celestijnenlaan 200F Chem&Tech 01.341, B-3001 Leuven tel: +32 16 327344 or +32 (0) 16 327142 (secr.) Home address Leuvensebaan 44, 3220 Holsbeek tel: +32 487 215951 E-mail work : [email protected] home: [email protected] Education 2001-2003 Bachelor in Chemistry, University of Leuven 2003-2005 Master in Biochemistry, University of Leuven (magna cum laude) Thesis title: Study of the interaction of HIV-1 integrase and LEDGF/p75 and characterization of monomeric red fluorescent proteins with fluorescence correlation spectroscopy (Promotor: Yves Engelborghs) 2005-2010 PhD in Science (IWT funded), option Biochemistry at Department of Chemistry, division Biochemistry, KU Leuven (promotors: Yves Engelborghs, Zeger Debyser). Thesis title: Confocal spectroscopy in living cells - Chromatin and protein interactions of HIV-1 integrase co-factor LEDGF/p75 Work history 2011-2013 Postdoctoral fellow of the Research Foundation Flanders (FWO), Department of Chemistry, division Molecular Imaging and Photonics, KU Leuven (promotor Johan Hofkens).

Transcript of 170228 Hendrix CV - Universiteit Hasselt Bioimaging Lab/1… · BOK protein multi-angled analysis...

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PersonaliaName JelleHendrixDay/Placeofbirth April28,1983,HasseltMaritalstatus marriedChildren twoUHasseltaddress OnderzoeksgroepFysiologie

VakgroepFysiologie-Biochemie-ImmunologieFaculteitGeneeskundeenLevenswetenschappenUniversiteitHasseltAgoralaanGebouwC–BIOMED,kantoorC034,B-3590Diepenbeek

tel:+3211269213KULeuvenaddress LaboratoryforPhotochemistryandSpectroscopy

DivisionofMolecularImagingandPhotonics DepartmentofChemistry Science,Engineering&TechnologyGroup KULeuven Celestijnenlaan200FChem&Tech01.341,B-3001Leuven tel:+3216327344or+32(0)16327142(secr.) Homeaddress Leuvensebaan44,3220Holsbeek tel:+32487215951E-mail work:[email protected]

home:[email protected]

Education2001-2003 BachelorinChemistry,UniversityofLeuven2003-2005 MasterinBiochemistry,UniversityofLeuven(magnacumlaude)

Thesis title: Study of the interaction ofHIV-1 integrase and LEDGF/p75and characterization of monomeric red fluorescent proteins withfluorescencecorrelationspectroscopy(Promotor:YvesEngelborghs)

2005-2010 PhD in Science (IWT funded), option Biochemistry at Department ofChemistry, division Biochemistry, KU Leuven (promotors: YvesEngelborghs,ZegerDebyser).Thesistitle:Confocalspectroscopyinlivingcells-ChromatinandproteininteractionsofHIV-1integraseco-factorLEDGF/p75

Workhistory2011-2013 Postdoctoral fellow of the Research Foundation Flanders (FWO),

DepartmentofChemistry,divisionMolecular ImagingandPhotonics,KULeuven(promotorJohanHofkens).

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2011-2013 Visiting scientist at Ludwig-Maximilians-University Munich, Germany(FWOfunded).

2013 Research fellow of the German Academic Exchange Service (DeutscherAkademischerAustauschdienst,DAAD).

2013-2016 Postdoctoral researcher (100% BAP) at Department of Chemistry,divisionMolecularImagingandPhotonics,KULeuven.

2014-2016 Lecturer(5%ZAP)atHasseltUniversity,DisciplinegroupChemistry.2014-2016 Coordinatorinteruniversity(Hasselt-Leuven)Bachelorinternships10-2016-… AssistantProfessor(tenuretrack,100%ZAP)‘Bioimaging’,UHasselt12-2016-… GuestProfessorshipKULeuven,MolecularImagingandPhotonics

Ongoingcollaborations1. Invitroandinvivostructuralandfunctionalanalysisofproteinsinvolvedinprotein

targetingandsorting.• Collaboration with KU Leuven - Rega institute – Molec. Bacteriology – Prof.

Anastassios Economou, Dr. Lily Karamanou and Marios Sardis.https://rega.kuleuven.be/bac/

2. Investigatingretrovirusesusingsingle-moleculemicroscopy• CollaborationwithKULeuven–Molec.VirologyandGeneTherapy–Prof.Zeger

Debyserandco-workers.http://www.kuleuven.be/molmed/3. StudyingconformationalswitchinginthemultidrugtransporterLmrP

• CollaborationwithULBBrussels–Prof.CédricGovaerts,Dr.ChloéMartensandAurélieRoth.http://govaertslab.ulb.ac.be

4. ConformationaldynamicsinelongationfactorTuduringtranslation• Collaboration with VIB-VUB Brussels - Prof. Remy Loris and Dr. Abel Pino

Garcia.http://www.vib.be/en/research/scientists/Pages/Remy-Loris-Lab.aspx5. Softwaredevelopmentforsingle-moleculespectroscopyanalysis

• Collaboration with Ludwig-Maximilians-Universität München – Prof. Don C.Lamb, Dipl. Chem. Waldemar Schrimpf and Dipl. Chem. Anders Barth.http://www.cup.uni-muenchen.de/pc/lamb/

6. Single-moleculeFRETonproteinsinvolvedinbacterialcellulosedegradation• Collaboration with Ludwig-Maximilians-Universität München – Prof. Don C.

Lamb.http://www.cup.uni-muenchen.de/pc/lamb/• CollaborationwithWeizmannInstitute,Rehovot,Israel–Prof.EdwardA.Bayer,

Dr. Daniel Fried and Dr. Yoav Barak.http://wws.weizmann.ac.il/Biological_Chemistry/scientist/Bayer/

7. Biomoleculardiffusioninbonemarrowandhydrogels• CollaborationwithKULeuven-Biomechanics–Prof.HansvanOosterwyck-

http://www.mech.kuleuven.be/en/bme/8. Studyofepidermalgrowthfactorusingfluctuationimaging

• CollaborationwithKULeuven-Biochemistry–LaboratoryforBiomolecularNetworkDynamics–Prof.HideakiMizuno–

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• http://chem.kuleuven.be/en/research/bmsb/bnd9. Fluorescentproteindevelopment

• Collaboration with KU Leuven – Biochemistry – Prof. Peter Dedecker -https://www.chem.kuleuven.be/pd/.

10. BOKproteinmulti-angledanalysis• CollaborationwithUNamur–Chemistry–Prof.CatherineMichaux.

Supervision- PhDpromotor:VeerleLemmens- PhDcopromotor:Dr.DoortjeBorrenberghs (IWT funded),DanaiLaskaratou (BOFfunded),BartVanHeertum(IWTfunded)

- PhD assessor: Niels Vandenberk (IWT funded), Milena Helmer-Lauer (joint PhDprogramme KU Leuven & Federal University of Espírito Santo), Jolien Blokken,SubhalakshmiSharma,OviaMargaret

- Master thesis promotor: Maarten Vervecken, Robbert Boudewijns, NielsVandenberk,DoortjeBorrenberghs,EvaMarting,GuillermoSolisFernandez,StevenMertens,DriesDavid,VeerleLemmens

- Masterthesismentor:AndersBarth(LMUMunchen)- Masterthesisreader:ElineBoons,LeenMathys,FloreDeWit,StefHoremans,DavidManuelRantasa

- Master internshippromotor:ThierryVerheyen,ErikSoons,MichaelVanhemelen,JorisRombouts,LievenLemaire,HerlindedeKeersmaecker

- Bachelorthesispromotor:EvaMarting(UHasselt),ChloeGeeroms,PieterNoyens,BeaTimmermans,HelenHamaekers,CharlotteDavid,LauranReyniers,Bert Jacobs,AlexMaes

- Bachelorthesismentor:SigurdVogler(LMUMunchen)- Bachelor thesis reader: Veerle van Meervelt, Hanne Dubois, Bert Roelants(KHLeuven),BartdeVos(KHLeuven),VeerleCrabbé(KHLeuven),NielsVandenberk(KHLeuven)

FundingPersonal- IWTPhDFellow of the Institute for the Promotion of Innovation through ScienceandTechnology in Flanders (IWTVlaanderen) fromOctober1st 2005– September30th2009.

- FWOpostdoctoralFellowoftheResearchFoundationFlanders(FWOVlaanderen)fromOctober1st2010–September30th2013.

- FWO travel grant for a long stay abroad (1 year) from Research FoundationFlanders(FWOVlaanderen)fromSeptember5th2011–August31st2012.

- DAAD Research Fellow of the German Academic Exchange Service (DeutscherAkademischerAustauschdienst,DAAD)fromMarch2013-June2013.

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- Travelgrantforattendingthe58thAnnualMeetingoftheBiophysicalSocietyinSanFrancisco,February14-19,2014fromtheBelgianSocietyforMicroscopy(BSM).

Grouplevel-granted- FWO research project (500k€, G0B4915N, 01/01/2015-31/12/2019) toLaboratoryofPhotochemistry&Spectroscopy

‘Development and application of broadly applicable microscopy methods andprobes for diffraction-unlimited fluctuation imaging of dynamic biologicalsystems.’January1,2015–December31,2018role:co-supervisor(supervisorJohanHofkens,co-supervisorLilyKaramanou),Idesignedandwroteproject

- Hercules large-scale infrastructure project (1.8M€, ZW15_09; 01/10/2016-31/09/2021) to a consortium (Dept. Chemistry, Centre for SurfaceChemistry andCatalysis,Dept.MechanicalEngineering)foracommerciallaserscanningmicroscopeequippedwith(amongotherthings)pulsedinterleavedexcitation.

Title:MultimodalfluorescencemicroscopyandnanoscopyplatformStatus:finaldecisionrole:co-promoter(promoterJohanHofkens,co-promotersHansvanOosterwyck,Peter Dedecker and Maarten Roeffaers). I had a substantial contribution inwrtingtheproject

- KULeuvenCat.1project(583k€,C14/16/053;01/09/2016-31/09/2020)totheLaboratoryforBiomolecularnetworkdynamics.

status:full-projectsubmission(passedLOIstage)role:co-supervisor(promoterHideakiMizuno,cosup.SusanaRocha),Idesignedthemethodologicalandpartofapplicationspartoftheproject.

- IWTscholarship(111595)toDoortjeBorrenberghs‘Developmentofanadvancedmodelsystemforstudyingretroviralreplicationatthesingle-viruslevel.’startdateJanuary1,2011role:promotor,Idesignedandcoordinateproject

- IWTscholarship(141515)toNielsVandenberk‘TheE.coliSecreactionpathwayforcellularproteinsortingunderaninnovativesingle-moleculeloupe’startdateJanuary1,2015role:assessor,Idesignedandcoordinateproject

- Scientific collaborator of KUL Cat. 1 project of Rik Schrijvers (C12/16/024,01/10/2016-31/09/2018)

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TeachingWith publication pressure being grafted onto every new scientist nowadays, theimportanceofhigh-leveleducationissometimesoverlooked.Luckily,however,Ihavea true passion for teaching science in a simple and understandable manner. In myopinion,thisistheonlywaypeople(scientistsornot)canappreciateitstruevalue.

Bachelorlevel- Former lecturerat theUniversityofHasselt, campusDiepenbeek(course: Inleidingtotdebiochemie(1399))–1semesterperyear,since2014

- CoordinatorinteruniversityinternshipsUHasselt-KULeuven- SpectroscopyofBiomolecules(B-KUL-G0O58C)(1lecture)- Geïntegreerdpracticum(B-KUL-G0O57C)(5years)- Immuniteit(3620)(Classonlabelsinimmunology)

Masterlevel- AdvancedEnzymology(B-KUL-G0U19A)(1lecture)- To become Coordinator of Advanced Fluorescence and FluorescenceMicroscopy(B-KUL-G0G59A)(2 lectures on fluorescence correlation spectroscopy, fluorescence recovery afterphotobleaching,singleparticletracking;4yearspracticals)

- AdvancesinMicroscopy(B-KUL-E08F8A)(1lecture)- LMU Munich (2011-2013) (3 lectures; instrumentation for microscopy, physicalbiochemistry,advancedfluorescencespectroscopy)

- ElectrophysiologyandImaging(1836)(multiplelectures)

PhDlevel- multiple(invited)lectures,seminarsandposterpresentations(+awards)- Istronglyencourageweeklyseminarsand‘journalclubs’,tobroadentheresearchers’minds.

- Speaker at 4 PhD student focused workshops and co-organizer of 4 PhD studentfocusedconferences.

Organizationofconferences- I helped in the organization of the International Conference of Photochemistry(ICP2013,Leuven,2013).

- I helped in the organization of the 3rd European Workshop on AdvancedFluorescenceImagingandDynamics(Munich,2012).

- Main organizer (together with Prof. Zeger Debyser) of a 1-day symposium onfluorescence imaging at the level of a single virus, in the context of theInteruniversity Attraction Pole BelVir IAP 7/45 ‘Virus-host interplay at the earlystagesofinfection’inLeuvenapril2016.

- Local organizing committee of the Methods and Applications in Fluorescence(MAF15)conferencein2017inBruges.

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EditorialactivitiesIreviewpapersonaregularbasisformanydifferentpeer-reviewedscientificjournals,suchasChemPhysChem,BiophysicalJournal,NucleicAcidsResearch,ScientificReports,CellResearch,FrontiersinMolecularNeuroscience,ChemComm...

Awards- Best poster presentation - LEDGF/p75 switches from a dynamic to a tightchromatininteractionuponbindingtoHIV-1integrase,TargetingHIVintegrationco-factors,2ndGeneralAssembly,Prague,CzechRepublic,29-30/01/09

- European Physical Journal award for best poster presentation - LEDGF/p75switches from a dynamic to a tight chromatin interaction upon binding to HIV-1integrase, Belgian Physical Society& Belgian Biophysical Society General ScientificMeeting,Hasselt,01/04/09

- ThirdprizeintheBelgianSocietyofMicroscopyPosterAward-Thetranscriptionalco-activatorLEDGF/p75displaysadynamicscan-and-lockmechanismforchromatintethering,AdvancedLightMicroscopySymposium,UniversityofGhent,23-24/09/10

ValorisationIn 2013 I tried to file an IP concerning the technology (PIE-FI microscopy, seeBibliography)Idevelopedwithmyco-workersatLMUMunich,butthetechnologywasnot patentable. Currently, we are collaborating directly with manufacturers (Zeiss,Leica), because making our development broadly accessible is important for thescientificcommunity.

Societalimpact- I strive forbroadlyaccessibleresearch,which is f.e.why Idirectly contacted thePress service of KU Leuven after the publication of our HIVwork ACS Nano. Thisresultedinthreebroad-audiencearticlesconcerningourwork:o Campuskrant, jaargang 25, nr. 9, 28/05/2014 – ‘Hiv-deeltje omgetoverd tot

proefbuis om geneesmiddelen te testen’(http://www.kuleuven.be/ck/files/ck25-nr09.pdf)(newspaperofKULeuven).

o Knackmagazine,nr.27,02/07/2014–‘Eiwittenvolgenineenvirus’(doorDirkDraulans) (http://magazine.knack.be/weekblad/magazines/02-07-2014/#,gedrukteversie)(KnackisaBelgian(Flemisch)weeklynewsmagazinecoveringlocalnews,politics,sports,business,jobs,andcommunityevents)

o Imaging&Microscopy Newsletter May 14, 2014 (http://www.imaging-git.com/news/single-molecule-fluorescence-imaging-new-technique-tracks-protein-single-hiv-particle)

- I gladly allow mentioning our work onmeta-scientific forums. For example, wewrote ameta-scientific review of our latestwork onHIV assembly in theAtlas ofScience: http://atlasofscience.org/on-their-way-out-structural-hiv-proteins-team-up-before-escaping-from-infected-cells/#more-1894.

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ResearchachievementsI am a Biophysicist who specializes in the development and application of single-molecule sensitivemulticolor fluorescencemicroscopymethods.With these, I aim atprovidingquantitativeinsightsincomplexbiologicalsystemsinvitroandinlivecells.

Expertise

BiochemistryIamknowledgeableinandhavepracticalexperiencewithmanyclassicalbiochemistrymethods such as DNA cloning, protein purification, gel chromatography, westernblotting, spectrophotometry/fluorometry, alpha-screen protein-protein interactionassay, chromatin fractionation assays, eukaryotic cell culture/transfection, virus/viralvector production and purification, ultracentrifugation, gradient centrifugation,multiparameterplatereading,ultrafiltration,dialysis...

OpticalmicroscopyI am an expert in fluorescence microscopy in general, and have much practicalexperience with wide-field fluorescence, confocal (laser scanning) microscopy, totalinternal reflection, single-molecule, ultrafast alternating excitation multicolormicroscopy(pulsedinterleavedexcitation),time-correlationsinglephotoncountingandFörsterresonanceenergytransfer.Iamalsoanexpertatbuildingsuchsystemsmyself.

BiologyI am very knowledgeable in the biology of HIV, and also work / have worked ondifferentaspectsoftranscription/translationmachineriesandmicrobiology.

Dataanalysis&programmingIamanexpertintimeorspacedependentcorrelationanalysisoffluorescencedataorimages. Specific methods include Fluorescence correlation spectroscopy and Rasterimage correlation spectroscopy, both in single and dual-color, and both withcontinuous-waveorpulsedexcitation/TCSPCdetection.Havingcollaboratedintimatelywithbiologistsoverthelast10years,Irealizetheimportanceofcomprehensive,easy-to-usesoftwareforrobustanalysisofallkindsoffluorescencedata.

Achievements

Whobindstowhominlivecells?Biochemicalsystemsofteninvolvetwoormultiplepartnersinteractingwitheachotherunder equilibrium conditions. Quantifying these equilibria in a live-cell approachprovidesmechanisticinsightsintothesesystems.AmajorresearchlineIputforwardduring my PhD was setting up fluorescence methods (e.g. FCCS) for quantifyingprotein-protein binding affinities in living cells. At the time, we were one of fewresearchgroupswhocoulddothisinaquantitativemanner.Iappliedthesemethodsinmy research on HIV-1 replication, resulting in multiple publications (De Rijck et al.,2006,Hombroucketal.,2007,Hendrixetal.,2011).Ialsocomplementedthesestudies

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withinvitroassaysforprobingPPIaffinities(McNeely,Hendrixetal.,2011).Mycurrentresearchcontinuesonthis.

FollowingproteinsinasinglevirusparticleOne of the objectives of my FWO postdoctoral project was engineering the humanimmunodeficiency virus such that protein-protein interactions could be studiedinsidesingleviralparticles,effectivelyturningthevirusintoa(safe)nanoscopictesttube.We firstpublishedadetailedmethodological studyon this (Borrenberghs et al.,2014).Beinginfectious,thisengineeredviruscould,however,alsobeusedtostudyHIVreplication. Inaseriesofotherstudies,wehaveused(Desimmieetal.,2012)andareusing(Borrenberghsetal.,twopapersinpreparation)thisprincipletolearnaboutHIVandmurine leukemiavirus.Thisworkhasbeencovered indifferentmedia (includingthenewspaperofKULeuvenandKnackmagazine).

AcolorfulfutureforfluorescenceimagingWithfundingfromFWOFlanders(Postdocgrantandtravelgrant),Idevelopedanewfluorescence imagingmethodat theuniversityofMunich (PIE-FI), that allowsanextraordinarily quantitative analysis of many biomolecular properties fromfluorescenceimagesobtainedbymulti-colorconfocalmicroscopy(Hendrixetal.,2012,Hendrixetal.,2013,Hendrixetal.,2014).PIE-FIupgradesaclassicalandwidelyusedconfocal microscope to an in vivo platform for biophysical chemistry. Althoughpublishedonlyin2013,thismethodisbeingcitedverywell,andwashighlightedintheBiophysicalJournalNewandNotablecolumn.Iampioneeringthis,andatKULeuvenIam currently setting up and further developing this novelmethod (IWT funded PhDprojectofNielsVandenberkandprojectfundingbyFWOn.G0B4915N).IalsorecentlypublishedadetailedinvestigationofHIVassemblywithPIE-FI(Hendrixetal.2015)andinacollaborativeeffort,PIE-FIwasusedtoprobeproteinsinvolvedinthemyelinsheetofneuronal cells (Ozgenet al., 2014). I like to refer toPIE-FI asnext-generation laserscanningmicroscopy.

ProbingbiomolecularfunctionbeyondstructureInlateyears,ithasbeenacceptedthatbiomolecularmacromoleculeshardlyeverformstatic 3D (tertiary) structures. Rather, a protein often structurally wobbles around,sometimesrandomly,butmoreoften thannotalso functionally.Such fuzziness isveryhard,ifnotimpossibletoinvestigateusingcommonlyusedstructuralbiologymethodssuchasX-rayorNMR.AnewresearchlinethatIsetupatKULeuvenisthequantitativeanalysisofproteinsuper-tertiarystructure(conformation)inrealtimeusingFörsterresonanceenergytransferatthesingle-moleculelevel.Thisisarelativelynewresearchfield thatperfectly complementsdynamic information frommolecularmodellingwithstaticinformationfrome.g.X-raystudies.Importantly,thebiophysicalresearchfieldinBelgium,thatencompassesmanystructuralbiologists,hasbeenawaitingamethodlikethis for a long time, I have come to notice. I am currentlywriting a paper aboutmy

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investigationsofabacterialcellulose-degradingenzymecomplexwiththisnewmethod,buthavepublishedabook chapteron this technology (Hendrix et al., 2011).Wealsorecentlypublishedadetailedinvestigationofaproteininvolvedinproteinfolding(Röhletal.,2015).DifferentnovelcollaborationsinBelgiumwillalsosoonleadtopublishableresults(seeOngoingcollaborationsandResearchPlan).

Attention,transcriptionfactorsatwork!AnothermajorgoalIputforwardduringmyPhDwasquantifyingchromatinbindingof transcription factors (TFs) in living cells with fluorescence correlationspectroscopy. In particular, I chose a TF implicated in the life cycle of the HIV,LEDGF/p75,toalsolearnaboutviralpathologyandkilltwobirdswithonestone.InthefirstpaperIwroteaboutthis(Hendrixetal.,2011),weprovidedadetailedinsightinthekineticregimethatdefinestheinsandoutsofLEDGF/p75intracellularmovement,andtheeffectofHIVproteinsonthis.ThispaperisverywellcitedintheHIVandchromatinfield. I also published some follow-up papers on this work, studying LEDGF/p75 indetailinacontrolledinvitroenvironment(McNeely,Hendrixetal.,2011andHendrix,VanHeertumetal.,2014),andstudyingother transcription factors(Bartholomeeusenetal.,2009)orHIVinhibitors(Desimmieetal.,2012)withthemethodologicaltoolboxIhadsetupduringmyPhD.

BetterprobesforfluorescencemicroscopyHigh-end fluorescencemicroscopyneeds thebest fluorescent probes. In our researchgroup,wedevelopneworganicdyebasedprobes, newnaturally fluorescentproteinswithtailoredproperties.Inoneparticularfirst-authorpublicationofmine(withresultsfrom my Master’s thesis)(Hendrix et al., 2008), I analyzed the spectroscopicpropertiesofsomenovelredFPs,andtheirperformanceinfluorescencecorrelationspectroscopy.Thispublicationisoneofmybest-citedpapers,sincecountlessresearchgroups are still using these probes. Today,we are still performing similar studies onotherprobes(near-infraredFPs,PhDprojectNielsVandenberk,Iammentor/assessor)and are developing novel RFPs with optimized quantum yields using a fluorescencelifetimescreeningapproach(FWOprojectn.G0B4915N,inwhichIamco-supervisor).

CompletepublicationlistSince 2006, I have published 21 articles in peer-reviewed internationally recognizedjournals, including top journals such as the Journal of Cell Biology (first author),ACSNano (last & corresponding author), Nucleic Acids Research (first author), NatureCommunications and PLoS Pathogens. I am corresponding author of 3 papers. I havepublished together with different groups at KU Leuven (Patrick Van Dijck, JohanThevelein, Zeger Debyser, Jan Delcour/Christoph Courtin, Guido Volckaert/RobLavigne),UGent(KevinBraeckmans/StefaanDeSmedt)andatuniversitiesworldwide(Germany,US,TheNetherlands).

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AccordingtoWebofSciencecitationreport‘HendrixJelle’onFebruary28,2017,thesecontributionshavebeencollectivelycitedover500times,withanaverageofabout21citationsperitem.Thishasresultedinacurrenth-indexof13:

Internationalpeer-reviewedjournals2017

• BurgerVM,VanderveldeA,Hendrix J,KonijnenbergA,SobottF,LorisR,StultzCM(2017).HiddenStateswithinDisorderedRegionsoftheCcdAAntitoxinProtein.JAmChem Soc. 2017 Feb 22;139(7):2693-2701. doi: 10.1021/jacs.6b11450 (IF 201613.93).

2016• Doortje Borrenberghs, Lieve Dirix, Flore De Wit, Susana Rocha, Jolien Blokken,

StéphanieDeHouwer,RikGijsbers, FraukeChrist, JohanHofkens, JelleHendrix*&Zeger Debyser* (2016). Dynamic Oligomerization of Integrase Orchestrates HIVNuclearEntry,ScientificReports6(doi:10.1038/srep36485)(*corresponding)

• Jelle Hendrix, Tomas Dekens, Waldemar Schrimpf and Don C. Lamb (2016).Arbitrary-Region Raster Image Correlation Spectroscopy, Biophysical Journal111(8):1785-1796(IFmostrecent:3.5).

2015• JelleHendrix,ViolaBaumgärtel,WaldemarSchrimpf,SergeyIvanchenko,MichelleA.

Digman, Enrico Gratton, Hans-Georg Kräusslich, BarbaraMüller and Don C. Lamb(2015).DirectobservationoftheonsetofHIV-1assemblyinlivingcells,TheJournalofCellBiology210(4):629-646(IFmostrecent:9.8).

• RöhlA,WenglerD,MadlT,LaglederS,TippelF,HerrmannM,HendrixJ,RichterK,Hack G, Schmid AB, Kessler H, Lamb DC, Buchner J (2015). Hsp90 regulates thedynamicsof itscochaperoneSti1andthe transferofHsp70betweenmodules.NatCommunications6:6655.(doi:10.1038/ncomms7655)(IFmostrecent:10.7)

2014

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• OzgenH.,SchrimpfW.,HendrixJ.,deJongeJ.C.,LambD.C.,HoekstraD.,BaronW.andKahya N. (2014) The Lateral Membrane Organization and Dynamics of MyelinProteinsPLPandMBPAreDictatedbyDistinctGalactolipidsand theExtracellularMatrix.PLoSONE9(7):e101834.(doi:10.1371/journal.pone.0101834)(mostrecentIF:3.73)(citations:1).

• Hendrix, J.*, Van Heertum, B.* and De Rijck, J. (2014) Dynamics of the ternarycomplex formedbyc-Myc interactor JPO2, transcriptional co-activatorLEDGF/p75and chromatin. Journal of Biological Chemistry 289(18):12494-506. (doi:10.1074/jbc.M113.525964)(mostrecentIF:4.65)(citations:1).*sharedauthorship

• Borrenberghs,D.,Thys,W.,Rocha,S.,Demeulemeester, J.,WeydertC.,Dedecker,P.,Debyser, Z., Hofkens, J., and Hendrix, J.* HIV virions as nanoscopic test tubes forprobing oligomerization of the integrase enzyme (2015).ACS Nano 8(4):3531-45.(doi:10.1021/nn406615v).(mostrecentIF:12.0).*correspondingauthoro Knackmagazine,nr.27,02/07/2014–‘Eiwittenvolgenineenvirus’(doorDirk

Draulans,gedrukteversie)(KnackisaBelgian(Flemisch)weeklynewsmagazinecoveringlocalnews,politics,sports,business,jobs,andcommunityevents)

o KU Leuven Campuskrant, jaargang 25, nr. 9, 28/05/2014 – ‘Hiv-deeltjeomgetoverd tot proefbuis om geneesmiddelen te testen’ (newspaper of KULeuven)

o Imaging&MicroscopyNewsletterMay14,2014o ScienceDaily

2013• Hendrix, J.*, Schrimpf, W., Holler, M., and Lamb, D.C.* (2013). Pulsed interleaved

excitation fluctuation imaging. Biophys J 105, 848-861. *corresponding author.(citations:11).(mostrecentIF:3.67).o HighlightedinNew&NotablecolumnoftheBiophysicalJournal:Wiseman,P.W.

(2013).FluctuationImagingSpicedUpwithaPieceofPIE.BiophysJ105,831.• Hendrix, J., and Lamb, D.C. (2013). Pulsed interleaved excitation: principles and

applications. Methods Enzymol 518, 205-243. (most recent IF: 2.00) (citations:8)(review).

• DeGraeve,S.,Marinelli, S., Stolz,F.,Hendrix, J.,Vandamme, J.,Engelborghs,Y.,VanDijck,P., andThevelein, J.M. (2013).Mammalian ribosomalandchaperoneproteinRPS3A counteracts alpha-synuclein aggregation and toxicity in a yeast modelsystem.BiochemJ455,295-306.(IFmostrecent:4.65)(citations:1).

• Desimmie,B.,Schrijvers,R.,Demeulemeester,J.,Borrenberghs,D.,Weydert,C.,Thys,W., Vets, S., Van Remoortel, B., Hofkens, J., De Rijck, J., Hendrix, J., Bannert, N.,Gijsbers, R., Christ, F., Debyser, Z. (2013). LEDGINs inhibit late stage HIV-1replicationbymodulatingintegrasemultimerizationinthevirions.Retrovirology,10,art.nr.10.1186/1742-4690-10-57,57.(citations:22)(mostrecentIF:5.66).

2012• Desimmie,B.,Humbert,M.,Lescrinier,E.,Hendrix,J.,Vets,S.,Gijsbers,R.,Ruprecht,

R., Dietrich, U., Debyser, Z., Christ, F. (2012). Phage display-directed discovery ofLEDGF/p75bindingcyclicpeptideinhibitorsofHIVreplication.MolecularTherapy,20 (11), art.nr. 10.1038/mt.2012.132, 2064-2075. (citations: 23) (IF publicationyear:7.04)(mostrecentIF:7.04).

2011

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• McNeely, M.*, Hendrix, J.*, Busschots, K., Boons, E., Deleersnijder, A., Gérard, M.,Christ, F., Debyser, Z. (2011). In vitro DNA tethering of HIV-1 integrase by thetranscriptional coactivatorLEDGF/p75. Journal ofMolecularBiology, 410 (5),811-830. (citations: 23) (IF publication year: 4.0) (most recent IF: 3.91). *sharedauthorship

• Cuyvers, S., Hendrix, J., Dornez, E., Engelborghs, Y., Delcour, J., Courtin, C. (2011).Both Substrate Hydrolysis and Secondary Substrate Binding Determine XylanaseMobilityasAssessedbyFRAP.JournalofPhysicalChemistryB,115(16),4810-4817.(citations:9)(IFpublicationyear:3.7)(mostrecentIF:3.61).

2010• Hendrix, J., Gijsbers, R., De Rijck, J., Voet, A., Hotta, J., McNeely, M., Hofkens, J.,

Debyser, Z., Engelborghs, Y. (2010). The transcriptional co-activator LEDGF/p75displaysadynamicscan-and-lockmechanismforchromatintethering.NucleicAcidsResearch,39(4),1310-1325.(citations:38)(IFpublicationyear:7.84)(mostrecentIF:8.28).

• Nath, S., Meuvis, J., Hendrix, J., Carl, S., Engelborghs, Y. (2010). Early aggregationsteps inalpha-synucleinasmeasuredbyFCSandFRET: evidence for a contagiousconformational change. Biophysical Journal, 98 (7), 1302-11. (citations: 42) (IFpublicationyear:4.22)(mostrecentIF:3.67).

• Ricicova,M., Kucharikova, S., Tournu, H., Hendrix, J., Bujdakova, H., Van Eldere, J.,Lagrou,K.,VanDijck,P.(2010).Candidaalbicansbiofilmformationinanewinvivoratmodel.Microbiology-SGM156,909-919.(citations:48)(IFpublicationyear:2.96)(mostrecentIF:2.85).

2009• Briers, Y., Schmelcher, M., Loessner, M., Hendrix, J., Engelborghs, Y., Volckaert, G.,

Lavigne,R.(2009).Thehigh-affinitypeptidoglycanbindingdomainofPseudomonasphageendolysinKZ144.BiochemicalandBiophysicalResearchCommunications,383(2),187-191.(citations:25)(IFpublicationyear:2.55)(mostrecentIF:2.41).

• Bartholomeeusen, K., Christ, F., Hendrix, J., Rain, J., Emiliani, S., Benarous, R.,Debyser, Z., Gijsbers, R., De Rijck, J. (2009). Lens epithelium-derived growthfactor/p75 interactswith the transposase-derivedDDEdomainofPogZ. JournalofBiological Chemistry 284 (17), 11467-11477. (citations: 50) (IF publication year:5.33)(mostrecentIF:4.65).

2008• Hendrix,J.,Flors,C.,Dedecker,P.,Hofkens,J.,Engelborghs,Y.(2008).Darkstatesin

monomericredfluorescentproteinsstudiedbyfluorescencecorrelationandsinglemolecule spectroscopy.Biophysical Journal, 94 (10), 4103-4113. (citations:84) (IFpublicationyear:4.68)(mostrecentIF:3.67).o HighlightedinBioPhotonics,anonlinejournaldiscussinglatestdevelopmentsin

thebiologicalapplicationsfieldofphotonics.• Buyens, K., Lucas, B., Raemdonck, K., Braeckmans, K., Vercammen, J., Hendrix, J.,

Engelborghs, Y., De Smedt, S., Sanders, N. (2008). A fast and sensitivemethod formeasuringtheintegrityofsiRNA-carriercomplexesinfullhumanserum.JournalofControlledRelease,126 (1),67-76. (citations:72) (IFpublicationyear:5.69) (mostrecentIF:7.63).

2007

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• Hombrouck,A.,DeRijck,J.,Hendrix,J.,Vandekerckhove,L.,Voet,A.,DeMaeyer,M.,Witvrouw, M., Engelborghs, Y., Christ, F., Gijsbers, R., Debyser, Z. (2007). VirusevolutionrevealsanexclusiveroleforLEDGF/p75inchromosomaltetheringofHIV.PLoSPathogens,3(3),e47.(citations:115)(IFpublicationyear:9.34)(mostrecentIF:8.14).

2006• DeRijck,J.,Vandekerckhove,L.,Gijsbers,R.,Hombrouck,A.,Hendrix,J.,Vercammen,

J., Engelborghs, Y., Christ, F., Debyser, Z. (2006). Overexpression of the lensepithelium-derived growth factor/p75 integrase binding domain inhibits humanimmunodeficiency virus replication. Journal of Virology, 80 (23), 11498-509.(citations:135)(IFpublicationyear:5.34)(mostrecentIF:5.08).

Inpreparation• Abel Pino Garcia, Hendrix J, Remy Loris – Role of phosphorylation in the

conformationofElongationfactorTu(submitted)• Borrenberghs D., Debyser Z., Hofkens J., Debyser Z., Hendrix J., Stoichiometry of

murineleukemiavirusintegraseenzymeinviralparticles(submitted)• Vandenberk N., Hofkens J., Hendrix J., Performance of FRET acceptors under PIE

conditions.• Vandenberk N., Hofkens J., Hendrix J., Fluctuation imaging using near-infrared

fluorescentproteins.• BarthA.*,HendrixJ.*,FriedD.*,BarakY.,Bayer,E.andLamb,D.C.FuzzinessofaC.

thermocellumscaffoldin conformation contributes to cellulosome function (* equalcontribution).

Internationallyrecognisedscientificbooks2014• Hendrix, J.* and Lamb, D.C.* (2014). Implementation and application of pulsed

interleaved excitation for dual-color FCS and RICS.Methods in Molecular Biology1076,653-682.*correspondingauthor(citations:4)

2011• Christ,F.,Busschots,K.,Hendrix,J.,McNeely,M.,Engelborghs,Y.,Debyser,Z.(2011).

Assays for evaluation of HIV-1 integrase enzymatic activity, DNA binding, andcofactorinteraction.In:NeamatiN.(Eds.),HIV-1Integrase.MechanismandInhibitorDesign,Chapt.12.Hoboken,NewJersey,USA:JohnWiley&Sons,Inc.,151-16

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Top-5publications‘GaugingGagassembly’HendrixJ,BaumgärtelV,SchrimpfW,IvanchenkoS,DigmanMA,GrattonE,KräusslichHG,MüllerB,LambDC.Live-cellobservationofcytosolicHIV-1assemblyonsetrevealsRNA-interactingGagoligomers.2015,JournalofCellBiology210(4):629-646(weblink).Journalimpactfactor:9.8(2014)Citedby:0(recentlypublished)

AbstractAssemblyoftheGagpolyproteinintonewviralparticlesininfectedcellsisacrucialstepintheretroviralreplicationcycle.Currently,littleisknownabouttheonsetofassemblyinthecytosol.Inthispaper,weanalyzedthecytosolicHIV-1Gagfractioninrealtimeinlive cells using advanced fluctuation imaging methods and thereby provide detailedinsights into the complex relationship between cytosolic Gagmobility, stoichiometry,andinteractions.WeshowthatGagdiffusesasamonomeronthesubsecondtimescalewithseverelyreducedmobility.Reductionofmobilityisassociatedwithbasicresiduesinitsnucleocapsid(NC)domain,whereascapsid(CA)andmatrix(MA)domainsdonotcontribute significantly. Strikingly, anotherdiffusiveGag specieswasobservedon theseconds timescale that oligomerized in a concentration-dependentmanner. Both NC-andCA-mediated interactionsstronglyassist thisprocess.OurresultsrevealpotentialnucleationstepsofcytosolicGagfractionsbeforemembrane-assistedGagassembly.Seealsothepopularizedabstract.

MotivationAlthoughalwaysmostproudofmylatestpaper,IfeelthisisthebestIhavepublishedsofar.Itcontainsthefourelementsthatreallydefinemeasaresearcher:

- Topical biological research question: We provided crucial insights in thecomplicatedHIVassemblysystem,whichuptillnow,hadbeenunattainable.

- Cutting-edge microscopy: We employ very powerful dual-color fluctuationimagingmethods,thatwe(recently)developedourselves.

- Advanced, freely available software:We helpmove forward the bio field bywritingeasy-to-usemicroscopyanalysisprograms(seeMIA).

- Comprehensive writing: The paper contains everything the reader needs tounderstandexactlyhowexperimentswerecarriedoutandinterpreted.

Furthermore, thisworkwas a successfulcollaborationbetweendifferent researchinstitutions, stationed worldwide (KU Leuven, UniversitätsKlinikum Heidelberg,Ludwig-Maximilians-UniversitätMünchenandUniversityofCaliforniaatIrvine).

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‘NewtechniquetracksproteinsinsingleHIVparticles’BorrenberghsD,ThysW,RochaS,DemeulemeesterJ,WeydertC,DedeckerP,HofkensJ,Debyser Z, and Hendrix J. HIV virions as nanoscopic test tubes for probingoligomerizationoftheintegraseenzyme.2014,ACSNano8(4):3531-3545(weblink).Journalimpactfactor:12.9(2014)Citedby:0(recentlypublished)

AbstractEmployingvirusesasnanoscopiclipid-envelopedtesttubesallowstheminiaturizationof protein-protein interaction (PPI) assays while preserving the physiologicalenvironmentnecessary forparticularbiologicalprocesses.Applied to thestudyof thehumanimmunodeficiencyvirustype1(HIV-1),viralbiologyandpathologycanalsobeinvestigated in novelways, both in vitro aswell as in infected cells. In thisworkwereportonanexperimentalstrategythatmakesuseofengineeredHIV-1viralparticles,toallowforprobingPPIsoftheHIV-1integrase(IN)insideviruseswithsingle-moleculeFörster resonance energy transfer (FRET) using fluorescent proteins (FP). We showthat infectious fluorescently labeled viruses can be obtained and that the quantity oflabels canbeaccuratelymeasuredandcontrolled inside individualviralparticles.Wedemonstrate,withpropercontrolexperiments,theformationofINoligomersinsingleviralparticlesandinsideviralcomplexesininfectedcells.Finally,weshowacleareffectonINoligomerizationofsmallmoleculeinhibitorsofinteractionsofINwithitsnaturalhumancofactorLEDGF/p75,corroboratingthatINoligomerenhancingdrugsareactivealready at the level of the virus and strongly suggesting the presence of a dynamic,enhanceableequilibriumbetweentheINdimerandtetramerinviralparticles.AlthoughappliedtotheHIV-1INenzyme,ourmethodologyforutilizingHIVvirionsasnanoscopictest tubes forprobingPPIs is generic, i.e., otherPPIs targeted into theHIV-1, orPPIstargetedintootherviruses,canpotentiallybestudiedwithasimilarstrategy.

MotivationThisismyfirstpaperaslast(corresponding)authorinLeuven;Idesignedtheproject(aspartofmyFWOpostdoc,1.2.702.11.N.00),carriedoutinitialexperiments,wrotethemanuscript andmanaged to get it into anexcellent journal. Importantly, duringmytwo-yearabsence foraresearchstay inMunich(FWOlongstayabroad,V444611N), Icontinuedpushingthisprojectforward.Tothebestofourknowledge,wewerethefirst tostudyprotein-proteininteractionsinside a single viral particle. Important biological insights were provided into HIV-1pathology/biology,and,anovelandverypowerfulandbroadlyapplicabletoolwasprovidedtostudyHIV-1orviruses ingeneral; inthemeantime,wepublishedanotherpaper using this method, and still two papers are in preparation (on HIV duringinfection, and on the murine leukemia virus). Because of its tangible character,differentmediawerealmost immediately interested inourwork(see f.e.KULeuvenCampuskrant,Knackmagazine27,July2nd,2104,Imaging&Microscopy).

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‘FluctuationimagingspicedupwithapieceofPIE’Hendrix J*, SchrimpfW,HöllerM,LambDC*.Pulsed interleavedexcitation fluctuationimaging.2013,BiophysicalJournal105(4):848-861(weblink).Journalimpactfactor:3.8(2013)Citedby:10

AbstractFluorescence fluctuation imaging is a powerful means to investigate dynamics,interactions, and stoichiometry of proteins inside living cells. Pulsed interleavedexcitation(PIE)isthemethodofnanosecondalternatingexcitationwithtime-resolveddetection andallows accurate, independent, andquasi-simultaneousdeterminationoffluorescence intensities and lifetimes of different fluorophores. In this work, wecombinepulsed interleavedexcitationwithfluctuation imagingmethods(PIE-FI)suchas raster image correlation spectroscopy (RICS) or number and brightness analysis(N&B). More specifically, we show that quantitative measurements of diffusion andmolecular brightness of Venus fluorescent protein (FP) can be performed in solutionwith PIE-RICS and compare PIE-RICS with single-point PIE-FCS measurements. Wediscuss the advantages of cross-talk free dual-color PIE-RICS and illustrate itsproficiency by quantitatively comparing two commonly used FP pairs for dual-colormicroscopy, eGFP/mCherry and mVenus/mCherry. For N&B analysis, we implementdead-timecorrectiontothePIE-FIdataanalysistoallowaccuratemolecularbrightnessdetermination with PIE-NB. We then use PIE-NB to investigate the effect of eGFPtandem oligomerization on the intracellularmaturation efficiency of the fluorophore.Finally, we explore the possibilities of using the available fluorescence lifetimeinformation in PIE-FI experiments, allowing us to quantitatively determinemolecularconcentrationsfrom100nMdownto<30pMwithPIE-rasterlifetimeimagecorrelationspectroscopy(RLICS).Weusethefluorescencelifetimeinformationtoperformarobustdual-color lifetime-based FRET analysis of tandem fluorescent protein dimers. Lastly,we investigate the use of dual-color RLICS to resolve codiffusing FRET species fromnon-FRETspeciesincells.Theenhancedcapabilitiesandquantitativeresultsprovidedby PIE-FImake it a powerfulmethod that is broadly applicable to a large number ofinterestingbiophysicalstudies.

MotivationThisisatoppublicationbecausebyinventingPIE-FI,Iveryelegantlysolvedproblemswithmulticolorfluorescenceimaging,thateveryoneintheworld(includingmyselfinLeuven!)wasexperiencing. Importantly, I specificallywent toMunich todevelop thismethod in theproperscientificenvironment(FWO longstayabroad,V444611N).Thereviewersofourpaperdefineditasoutstanding,sotheBiophysicalJournalevenwroteaNew&Notablecolumnaboutit.ToadvertisePIE-FI,andtoestablishmyselfasexpertin photon counting confocalmicroscopy, I became co-corresponding (*) author onthis paper, and wrote chapters in the new bible for Fluorescence FluctuationSpectroscopyand ina fluorescencebook inMethods inMolecularBiology.PIE-FIwasalso included in the book covering the latest top-notch developments in Advanced

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PhotonCounting.TogetherwithLeica(non-disclosureagreement),wearecurrentlyco-developingcommercialplatformswiththismethodology.

Attention,transcriptionfactorsatwork!Hendrix J, Gijsbers R, De Rijck J, Voet A, Hotta J, McNeely M, Hofkens J, Debyser Z,Engelborghs Y. The transcriptional co-activator LEDGF/p75 displays a dynamic scan-and-lockmechanismforchromatintethering.2011,NucleicAcidsResearch39(4):1310-25(weblink).Journalimpactfactor:8.0(2011)Citedby:32

AbstractNearlyallcellularanddiseaserelatedfunctionsofthetranscriptionalco-activatorlensepithelium-derived growth factor (LEDGF/p75) involve tethering of interactionpartners to chromatin via its conserved integrase binding domain (IBD), but little isknownabout themechanismof invivochromatinbindingand tethering. In thisworkwestudiedLEDGF/p75inreal-timeinlivingHeLacellscombiningdifferentquantitativefluorescencetechniques:spotfluorescencerecoveryafterphotobleaching(sFRAP)andhalf-nucleus fluorescence recovery after photobleaching (hnFRAP), continuousphotobleaching, fluorescence correlation spectroscopy (FCS) and an improved FCSmethod to study diffusion dependence of chromatin binding, tunable focus FCS.LEDGF/p75movesaboutinnucleioflivingcellsinachromatinhopping/scanningmodetypicalfortranscriptionfactors.ThePWWPdomainofLEDGF/p75isnecessary,butnotsufficient for in vivo chromatinbinding.After interactionwithHIV-1 integrase via itsIBD,ageneralprotein-proteininteractionmotif,kineticsofLEDGF/p75shiftto75-foldlarger affinity for chromatin. The PWWP is crucial for locking the complex onchromatin. We propose a scan-and-lock model for LEDGF/p75, unifying paradoxicalnotionsoftranscriptionalco-activationandlentiviralintegrationtargeting.

MotivationInwhat I call thecrownonmyPhDwork, I show for the first timenot being shyofadvancedfluorescencemethods,andofusingthemtogetthemaximuminformationoutofcomplexbiologicalsystems.Importantly,theapproachfollowedprovesthatthesumismorethanitsparts; interdisciplinaryresearch(inthiscase,virology&biophysics)allows providing answers to research questions that are unattainable otherwise. Ipracticethismodeofthoughttothisday.Nexttothis,eventhoughIonlywasaPhDstudentatthetime,Idesignedthisresearchproject myself, and single-handedly set up in Leuven advanced microscopymethods, thatpeoplearestillusing to thisday.At the time, thispaperwas themostquantitative investigation by fluorescencemicroscopy that had been carried out inourresearchgroup(Prof.Em.YvesEngelborghs).Also,Ideliberatelyaimedforahigherimpact journal thanwas average in the group, to reach outmuch furtherwithmyresearch. This work is very well-respected in the biophysics field of quantitativeimagingofchromatinbindingkinetics.

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Becarefulwhenanalyzingredfluorescentproteins!HendrixJ,Flors,C,DedeckerP,HofkensJ,EngelborghsY.Darkstatesinmonomericredfluorescent proteins studied by fluorescence correlation and single moleculespectroscopy.2008,BiophysicalJournal94(10):4103-4113(weblink).Journalimpactfactor:4.9(2008)Citedby:68

AbstractMonomeric red fluorescent proteins (mRFPs) have become indispensable tools forstudyingproteindynamics,interactionsandfunctionsinthecellularenvironment.Theiremission spectrum can be well separated from other fluorescent proteins, and theirmonomeric structure preserves the natural function of fusion proteins. However,previousphotophysicalstudiesofsomeRFPshaveshownthepresenceoflight-induceddarkstatesthatcancomplicatetheinterpretationofcellularexperiments.Inthisarticle,we extend these studies to mRFP1, mCherry, and mStrawberry by means offluorescence correlation spectroscopy and prove that this light-driven intensityflickering also occurs in these proteins. Furthermore, we show that the flickering inthese proteins is pH-dependent. Single molecule spectroscopy revealed reversibletransitionsfromabrighttoadarkstateinseveraltimescales,evenuptoseconds.Time-resolvedfluorescencespectroscopyshowedmultiexponentialdecays,consistentwitha"loose'' conformation.Weoffera structuralbasis for the fluorescence flickeringusingknowncrystal structuresandpointout that theenvironmentofGlu-215 iscritical forthe pH dependence of the flickering in RFPs. We apply dual-color fluorescencecorrelation spectroscopy inside live cells to prove that this flickering can seriouslyhampercellularmeasurements if thetimescalesof the flickeringanddiffusionarenotwellseparated.

MotivationThis ismy first first-author paper that I wrote based on experiments I carried outduringmyMaster’s thesis. Itwas the first in-depth analysis ofmRFPs,which at thetimewerebecomingthe fluorescent labelsofchoice formulticolor imaging inside livecells.Anexampleofdoingtherightsetofexperimentsattherighttime,itbecameawell-respected and verywell-cited work in the fluorescence field. It still receivesabout 10 citationsper year. Itwas also referenced in Biophotonics, an online journaldiscussinglatestdevelopmentsinthebiologicalapplicationsfieldofphotonics.

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Studiesabroad- Iworkedfortwoyears(9/2011–8/2013)asavisitingscientistattheuniversityofMunich(Ludwig-MaximiliansUniversität,oneof11ExcellenzInitiativuniversitiesinGermany)withProf.DonC.Lamb,withtheaimofbringingcutting-edgeexpertiseinadvancedfluorescencemicroscopytoFlanders/Belgium.Isetupnovelfluorescencemethods in Munich, and carried out multiple investigations of complex biologicalsystems(seeTop-5publications).IcontinuethecollaborationwithProf.Lambtothisday.

- Duringmy PhD, in 2007, I attended a 1-week fluorescence spectroscopy course atPicoquant,Berlin.

ConferencesI regularly attend the most important national and international meetings in thebiophysics field,where I always represent ouralmamaterwith a lecture (sometimesposter). Putting the KU Leuven on the globalmap is as important as doing research,publishingandteaching.

Invitedtalks- VisualizingandquantifyingHIV-hostinteractionswithfluorescencemicroscopy,8thEuropeanBiophysicsCongress,Budapest,Hungary,23-27/8/11

- Probing biomolecular structure and function withsingle-photon sensitivefluorescence imaging, Modern Biophysical Techniques for the Life Sciences,KoninklijkeAkademievoorWetenschap,Brussel,21/10/14

- Keynote lecture - Physics on the Scale of the Cell – Theoretical concepts andExperimental Methods, September 28 – October 2, 2014, University of Bayreuth,Germany.

- NIMWorkshoponAdvancedFluorescenceMethods,December8-12,2014inMunich,Germany.

- Modern biophysical methods for protein-ligand interactions, 1–5 June 2015, Oulu,Finland

- Research seminar,Medical School Leuven, October 2, 2015 (invited by Prof. ZegerDebyser).

- NIMWorkshoponAdvancedFluorescenceMethods,December5-19,2016inMunich,Germany.

- 2nd Hands-on-Light Microscopy Workshop, February 20-24, 2017, University ofLuxembourg(InvitedbyZeissMicroscopy).

Internationaltalks,abstract-selected- Targeting replication and integration ofHIV - TRIoHGeneral Assembly, Barcelona,Spain,8-12/12/06

- Joint Biophysical Society 52nd Annual Meeting and 16th IUPAB InternationalBiophysicsCongress,LongBeach,California,USA,2-6/02/08

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- TargetingHIVintegrationco-factors-2ndGeneralAssembly,Prague,CzechRepublic,29-30/01/09

- 7thEuropeanBiophysicsCongress,Genoa,Italy,11-15/07/09- 12thCarlZeisssponsoredworkshoponFCSandrelatedmethods,Cargèse,Corsica,12-16/10/09

- 18th International Workshop on Single Molecule Spectroscopy and UltrasensitiveAnalysisintheLifeSciences,Berlin,Germany,5-7/9/12

- 3rdEuropeanWorkshoponAdvancedFluorescenceImagingandDynamics,October8-12,2012inMunich,Germany

- Dutch meeting on Molecular and Cellular Biophysics, Veldhoven, The Netherlands30/9-1/10,2013

- 26thInternationalConferenceonPhotochemistry,July21-26,2013,Leuven,Belgium- BiophysicalSociety57thAnnualMeeting,February2-6,2013inPhiladelphia,USA

Internationalposterpresentations,abstract-selectedForposterprizes,see‘Awards’- Targeting replication and integration ofHIV - TRIoHGeneral Assembly, Barcelona,Spain,8-12/12/06

- Zeiss International Workshop on Fluorescence Correlation Spectroscopy methods,Stockholm,Sweden,4-6/12/06

- Joint Biophysical Society 52nd Annual Meeting and 16th IUPAB InternationalBiophysicsCongress,LongBeach,California,USA,2-6/02/08

- 7th InternationalWeber Symposium on Innovative FluorescenceMethodologies inBiochemistry and Medicine and 11th International Workshop on FluorescenceCorrelationSpectroscopyandRelatedMethods,Kauai,Hawaii,USA,6-12/06/08

- 58thAnnualMeetingoftheBiophysicalSociety,SanFrancisco,15-19February2014- Nationalposterpresentations,abstract-selected- I attended multiple local symposia and conferences where I presented posters,withoutabstractselection.

- Belgian Physical Society & Belgian Biophysical Society General Scientific Meeting,UniversiteitHasselt,01/04/09

- AdvancedLightMicroscopySymposium,UniversityofGhent,23-24/09/10

OthercompetencesLanguage- Dutch,English:nativeo I attended the Academic English course of the Arenberg Doctoral School,

althoughmyexperiencecomesmostlyfromwritingpapersandgrantproposals.- German,French:B2- Spanish,Russian:A2

Computer- Mac&windows:advanced- Word,Excel,Onenote,Powerpoint,Coreldraw,Origin:expert

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- MATLAB(programming):expert- IgorPro(programming):beginner

Leadershipskills- Leadingaresearchteam–4daycourse

Recruitment- In2007, Ideveloped(togetherwithWernerVerbakel)a ‘crimescene investigation’do-it-yourselfexperimentfortheinfodaysofBiochemistryatKULeuven.Thisisstillbeingused.

- IactivelyscoutfortalentedChemistry,Biochemistry,BioinformaticsandBiophysicsstudentstojoinourresearchgroups.