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SriramSubramaniamHong-WeiGaryblue:actin

Ciminietal.,2006,DynamicHong-Wei微管的組成成分+80

OnlyGTP-boundtubulinGTPishydrolyzedAGTP-tubulincapstabilizesmicrotubuleplusendGDP-tubulintendstodisassemblebutisconstrainedwithinthemicrotubulelatticeAconformationalchangeoccursuponlossofGTP-capthatresultsindisassembly

Nogalesetal.1998Nature;L?weetal.,2001JMBHong-WeiHong-WeiAThreeMinutesIntroductionofHong-WeiScratchoftheideaofTEM(March9,1931)FromtheNobelLecturebyErnst

FirstTEMbyErnstRuskaFrom

ErnstRuska(1906–NobelPrizeinPhysics“forhisfundamentalworkinelectronoptics,andforthedesignofthefirstelectronmicroscope”Hong-Wei第一 的電子顯微Thevisualizationof intheHelmantRuska(1908–1973)

Thefirststructuralinformationabouta TMVisafilamentous Hong-Wei

Hong-WeiTransmissionelectronmicroscopesErnstRuska, JEOL300KVHong-Wei ai ai300kVTitanHong-Wei電子顯微 物樣 的問(wèn)MakingthespecimenthinenoughtogetanelectronbeamthroughPreservingandcontrastingthevariousstructuralelementsofthematerialthehighvacuumofthemicroscope.(GlaeserRMandTaylorK,JSB,Problems:?AbhortheLowIntrinsicRadiationConventionalTEMofFixation,embedding,ultra-sectioning,and

stainstainproteinsandnucleicHong-Wei細(xì)胞超薄切片的電子顯 :10Fig.1-HelenHong-WeiMyosin Metal

FrozenFig.6-Bartesaghietal.,Science3Dreconstructionof-Galactosidaseusingsingleparticleat2.2Angstrom

Hong-Wei冷凍電鏡解 的高分辨率結(jié) Zhangetal.,2010,Hong-Wei2Dcrystalsofmembraneproteinsreachhighatomicresolution:1.8Gonenetal.,NatureHong-WeiTRPV1constructusedinthisworkisatetramerwithMWof~300kDa,purifiedfromHEK293cells.Liaoetal.,NatureHong-WeiTRPV1@3.4Liaoetal.,Nature-secretase@4.5Luetal.,Nature

Complex1@4.9Vinothkumaretal.,Nature

RyR-1@4.8Zalketal.,Nature2015;RyR-1@3.8?Yanetal.,NatureTRPA1@4.0Paulsenetal.,Nature

Anthraxtoxin@2.9?Jiangetal.,Nature2015

Hong-WeiHong-WeiHong-WeiHong-Wei ysisfrommoleculestoRibosome(3 (~120 Eukaryotic

TRPV1(300Hong-Wei ysisofsamplesnotsuitableforX-raycrystallographyand

Lietal.,

ActinfilamentRouilleretal.,Hong-WeiElectronTomography（電子斷層掃描三維重構(gòu)McIntoshetal.,Hong-WeiFromJensen’sLabatCalHong-WeiStructuraldeterminationinsolution,avoidingtime-consumingcrystallizationprocessandcrystallographicartifact.Candeterminenanometertosub-nanometerresolutionstructureswithverylittleamountofsample3μlof0.14,0002nmHong-Wei20μlof0.1μM(~600kDa)frozensampleshippedfromUStoTsinghua.3μlofsolutionononegood200micrographstakenonTitanKriosmicroscopein5~20,000particleimagestogeta6.5AngstromresolutionHong-WeiSlidefromTomWalz’s2009ScrippsworkshopHong-WeiScheresetal.2007,NatHong-Wei1.5millionparticledatasetclassifiedintomorethan30groupsetal.2010,Hong-WeiHowtocorrelateinvitrostructuralinformationtotheinvivoFEI

LowMagHighMagEMSartorietal.,2007,J.Struct.Phasecontrast Fluorescence

Hong-WeiAutomationofCourtesyofBridgetHong-Weiblue:actinGaryDr.TorstenSkeletalCampelloneetal.,2008,Cargotransport

Ciminietal.,2006,DynamicHong-WeiLow Chemicalenergyenergy

energyenergystored

energycharge

Hong-WeiConlyHong-WeiHong-Wei著絲粒復(fù)合體Dam1Theyeast

Dam1complex Westermannetal.2005Mol.CellMirandaetal.2005NMSBHong-WeiDam12-Helicalassemblyissensitiveto

300

Wangetal.2007

600

Hong-WeiDam113-fold16-fold

150

80

2-300600

Hong-Wei解聚微管可以推動(dòng)Dam1Dam1complexformsringsaroundmicrotubulesWestermannetal.2005Mol.CellWestermannetal.2006NatureHong-WeiKinesin:5Grinetal.2005Mol.Mirandaetal.2005DepolymerizingMTisamolecularTheenergycomesfromGTPthatisconsumedduringtheMTThecurlingprotofilamentsservepower-strokestodrivetheHong-WeiHong-WeiWhittakeretal.,1995,Hong-WeiWalkeretal.,2000,Post- Dominguezetal.,1998, RonHong-WeiHong-WeiYon

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