相关文章
促销商品
最新问答
- [07-28]怎么样正确的进行ELISA试剂盒的检测
- [07-25]哪家公司的ELISA试剂盒质量比较好
- [10-03]看过来!世界著名抗体公司汇总全在这里2017
- [10-03]怎么看一个公司代测的ELISA试剂盒准不准_问答详情_蛋白纯化_...
- [08-10]如何选择合适的流式抗体(五)优宁维(univ)官网
- [10-03]用于提高肺炎衣原体抗原/支原体抗原在免疫层析试剂中稳定性的包被...
- [10-03]双抗夹心法制备Elisa试剂盒时。怎么确定包被用抗体浓度?
- [07-26]纯化抗体
- [08-13]【求助】DNA甲基化纯化试剂盒的购买 实验方法
精选品牌
热卖商品
Crystallization of Kinesin Family Motor Proteins
蚂蚁淘
/发布时间:1545758271 /浏览量:195
| Motorproteinsofseveralkinesinfamilygroupshavenowbeencrystallized:monomericKinesin-1motordomainsfromhuman,ratandNeurospora(Kulletal.,1996;Sacketal.,1997;Songetal.,2001)anddimericratKinesin-1(twomotordomainsconnectedbyashortcoiled-coil;Kozielskietal.,1997);theKinesin-14(formerlyC-terminalmotor)proteinsDrosophilaNcdinitsmonomericanddimericforms(Sablin&Fletterick,1995,Sablinetal.,1996,1998;Kozielskietal.,1997;Yunetal.,2003),yeastKAR3(monomer)(Gulicketal.,1998;Yunetal.,2001)andthreeKAR3mutants(Yunetal.,2001),andKCBP(Vinogradovaetal.,2004);theKinesin-3(formerlyUnc104/KIF1)motor,KIF1Aboundtodifferentnucleotides(KIF1A-ADP,KIF1A-AMPPCP,KIF1A-ADP-Vi,KIF1A-AlFx)(Kikkawaetal.,2001;Nittaetal.,2004);theKinesin-5(formerlyBimC)motor,monomericEg5(Turneretal.,2001);andtheKinesin-13(formerlyMCAK)motor,PfKinI(Shipleyetal.,2004).Thefollowingtablessummarizethecrystallizationconditionsandsomeofthecrystalparameters. |
 |
| CrystalsofmonomericratKinesin-1 |
| Construct | Method | Conditions | References |
| HumanKinesin-1hK349 | Sittingdrop4°C | 5mg/mlproteinin50mMNa-acetate,pH4.6,75mMKCl,3.5%(w/v)PEG4000,2.5mMATP,10mMMgCl2Reservoir:100mMNa-acetatepH4.6,150mMKCl,7%PEG4000,5mMATP,20mMMgCl2 | Kulletal.,1996 |
| Ncd335-700 | SittingdropRoomtemperature | 7mg/mlproteinin10mMPipes,pH7.2,100mMNaCl,1mMEGTA,1mMDTT,7%(w/v)PEG4000,0.3%octyl-ß-D-glucoside,2mMATP,10mMMgCl2Reservoir:15%(w/v)PEG4000,60mMNaClequallybuffered | Sablin&Fletterick,1995Sablinetal.,1996 |
| Ncd293-700 | Hangingdrop18°C | 17mg/mlproteinin20mMHEPESpH7.4,200mMNaCl,10mMMgCl2,2mMDTTwaspre-incubatedwith4mMAMP*PNPorATPfor2hoursCrystalsgrewin11%PEG8000,0.8MNaCl,50mMNa2H2PO4,pH6.8,7mMDTT | Yunetal.,2003 |
| RatKinesin-1rK354 | HangingdropRoomtemperature | 9-14mg/mlprotein20mMPIPESpH7.5,50mMKCl,1mMEGTA,1mMDTT,0.9MLi2SO4Reservoir:20mMPIPESpH7.5,1.8MLi2SO4or15mg/mlproteinin10mMPIPESpH7.5,50mMNaCl,1mMEGTA,1mMDTT,0.5mMNaN3,0.9M(NH4)2SO4Reservoir:1.8M(NH4)2SO4,50mMNaCl | Kozielskietal.,1997aSacketal.,1997 |
| RatKinesin-1rK379 | HangingdropRoomtemperature | 15mg/mlprotein10mMPIPESpH7.5,200mMNaCl,1mMEGTA,1mMDTT,0.5mMNaN3,0.8M(NH4)2SO4Reservoir:1.6M(NH4)2SO4,200mMNaCl | Kozielskietal.1997a,b |
| Kar3383-729 | MicrobatchRoomtemperature | 11mg/mlprotein10mMHEPESpH7.5,150mMNaCl,1mMDTT,1mMMgCl2,0.2mMNaN3,2mMADPcombined1:1with22%methyletherPEG2000,100mMNaCl,2%ethyleneglycol,50mMHEPESpH7.0 | Gulicketal.,1998 |
| Ncd281-700 | Sittingdrop4°C | 20mg/mlproteinin20mMHEPESpH7.5,100mMNaCl,1mMEGTA,0.7MLi2S04,2mMADP,10mMMgCl2Reservoir:20mMHEPESpH7.5,1.4MLi2SO4,1mMEGTA,1mMDTT,10mMMgCl2 | Sablinetal.,1998 |
| Ncd295-700 | HangingdropRoomtemperature | 5mg/mlproteinin25mMNa2PO4,pH6.8,6.8%PEG8000,1MNaCl,2mMATP,3.5mMDTT,10mMMgCl2Reservoir:25mMNa2PO4pH6.8,13.5%PEG8000,2MNaCl,7mMDTT | Kozielskietal.,1999 |
| Eg51-368HsKSP | SittingDrop4°C | 5mg/mlproteinin9%PEG-3350,50mMPIPESpH6.8,100mMNaNO3Reservoir:18%PEG-3350,100mMPIPESpH6.8,200mMNaNO3Imperfectcrystalswerecrushedandusedtoseed5mg/mlEg5in7.5%PEG-3350,50mMMESpH5.6,100mMNaNO3Reservoir:15%PEG-3350,100mMMESpH5.6,200mMNaNO3 | Turneretal.,2001 |
| KIF1A-ADP | Vapordiffusion | 2microlitersofprotein(15mg/ml)+2microlitersofreservoirbuffer(RB1)composedof30%w/vPEG4000,100mMTris-HClpH8.5,200mMsodiumacetate,8%w/vsucrose,4mMADP,10mMMgCl2,equilibratedagainstRB1for5d | Kikkawaetal.,2001 |
| KIF1A-AMPPCP(soaked) | Vapordiffusion | 15mg/mlprotein+reservoirbuffer(RB2)composedof27%w/vPEG4000,100mMMES-NaOHpH6.5,200mMsodiumacetate,thensoakedinRB2+20mMAMPPCP+40mMMgCl2for24hrs. | Kikkawaetal.,2001 |
| KIF1A-AMPPCP(co-crystalized) | Vapordiffusion | 15mg/mlproteinco-crystalizedwith5mMAMPPCP+20mMMgCl2inreservoirbuffer(RB3)composedof30%w/vPEG4000,100mMTris-HClpH8.5,200mMsodiumacetate | Kikkawaetal.,2001 |
| Nkin1-355 | SittingDrop19°C | 7.5-15mg/mlproteinin20mMTrispH7.9,5mMMgCl2,0.5mMADPReservoir:17.5%PEGMME2000,100mMHEPESpH6.5-7.5,3%glycerol | Songetal.,2001 |
| Kar3+N11(WT) | HangingDrop18°C | 2microlitersofprotein(15mg/ml)+2microlitersofwellsolutioncontaining20-26%PEG2000ME,0.2MNaCl,50mMHEPESbufferpH7.0-8.0 | Yunetal.,2001 |
| Kar3N650K | HangingDrop18°C | 2microlitersofprotein(15mg/ml)+2microlitersofwellsolutioncontaining20-26%PEG2000ME,0.2MNaCl,50mMHEPESbufferpH7.0-8.0 | Yunetal.,2001 |
| Kar3R598A | HangingDrop4°C | 2microlitersofprotein(15mg/ml)+2microlitersofwellsolutioncontaining20-26%PEG2000ME,0.2MNaCl,50mMHEPESbufferpH7.0-8.0 | Yunetal.,2001 |
| Kar3E631A | HangingDrop4°C | 2microlitersofprotein(15mg/ml)+2microlitersofwellsolutioncontaining20-26%PEG2000ME,0.2MNaCl,50mMHEPESbufferpH7.0-8.0 | Yunetal.,2001 |
| Ncd293-700 | 18°C | 17mg/mlproteinin20mMHEPESpH7.4,200mMNaCl,10mMMgCl2and2mMDTT,pre-incubatedwith4mMAMPPNPorATPfor2hCrystalsgrewin11.0%PEG8000,0.8MNaCl,50mMNa2HPO4/NaH2PO4(pH6.8)and7mMDTTat18°C | Yunetal.,2003 |
| KIF1A | Vapordiffusionat20°Cfor24h | AMPPNP:27%w/vPEG4000,100mMTris-HClpH8.5,200mMsodiumacetateand3%w/vxylitolwithafinalconcentrationof5mMAMPPNPand1mMMgCl2ADP-AlFx:29%w/vPEG4000,100mMTris-HClpH8.5,200mMsodiumacetateand3%w/vxylitolwithafinalconcentrationof1mMADP,1mMMgCl2and1mMAlF3ADP-Vi;28%w/vPEG4000,100mMTris-HClpH8.5,200mMsodiumacetateand3%w/vxylitolwithafinalconcentrationof1mMADP,1mMMgCl2and1mMNaVO4 | Nittaetal.,2004 |
| KCBP884-1252 | Sittingdropvapordiffusionat4°C | 10mg/mLproteinin50mMTris,pH7.5,50mMNaCl,2mMMgCl2,1mMEGTA,1mMATP,1mMTris(2-carboxyethyl)-phosphineReservoir:20%polyethyleneglycol3350in0.2Mdi-sodiumhydrogenphosphate,pH9.1 | Vinogradovaetal.,2004 |
| KinI | Sittingdrop4°C | 10-20mg/mlproteinReservoir:1.4-1.8Mammoniumsulfate,100mMsodiumacetate(pH5.0),200mMsodiumnitrate | Shipleyetal.,2004 |
| Construct | Spacegroup | Unitcell | Resolution | Structuraldetermination | SpecialFeatures |
| hK349PDB:1BG2 | P212121 | a=48.54Åb=67.94Åc=112.95Å | 1.8Å | Multipleisomorphousreplacement | |
| rK354PDB:2KIN | P212121 | a=71.56Åb=73.67Åc=74.13Å | 1.9Å | Molecularreplacement | |
| rK379PDB:3KIN | P212121 | a=72.2Åb=91.9Åc=141.7Å | 3.0Å | Multipleisomorphousreplacement | |
| Kar3382-729PDB:3KAR | P21 | a=44.1Åb=81.2Åc=48.3Åß=105.8° | 2.3Å | Molecularreplacementandphasesofthreeheavyatomderivatives | |
| Ncd335-700Coordinates | I222 | a=127.1Åb=122.3Åc=68.0Å | 2.5Å | Multipleisomorphousreplacement | |
| Ncd281-700PDB:2NCD | P6122 | a=123.0Åb=123.0Åc=121.1Å | 2.5Å | Molecularreplacement | |
| Ncd295-700PDB:1CZ7 | C2221 | a=116.19Åb=148.83Åc=261.52Å | 2.9Å | Molecularreplacementandphasesofthreeheavyatomderivatives | |
| Eg5PDB:1II6 | P21 | a=53.08Åb=78.59Åc=94.15Åß=93.84° | 2.1Å | Molecularreplacement | |
| KIF1A-ADPPDB:1I5S | P212121 | a=41.67Åb=51.92Åc=157.06Å | 2.2Å | Molecularreplacement | |
| KIF1A-AMPPCP(soaked)PDB:1I6I | P212121 | a=41.99Åb=56.40Åc=156.12Å | 2.0Å | Molecularreplacement | MotorboundtoAMPPCP |
| KIF1A-AMPPCP(co-crystalized)PDB:1IA0 | P212121 | a=42.42Åb=55.43Åc=157.27Å | 1.9Å | Molecularreplacement | MotorboundtoAMPPCP |
| Nkin1-355PDB:1GOJ | P212121 | a=51.97Åb=72.73Åc=84.93Å | 2.3Å | Molecularreplacement | |
| Kar3+N11(WT)PDB:1F9T | P21 | a=43.6Åb=78.8Åc=47.2Åß=105.0° | 1.5Å | Molecularreplacement | |
| Kar3N650KPDB:1F9U | P21 | a=43.6Åb=78.0Åc=47.3Åß=105.1° | 1.7Å | Molecularreplacement | |
| Kar3R598APDB:1F9V | P21 | a=43.9Åb=77.4Åc=47.7Åß=105.9° | 1.3Å | Molecularreplacement | |
| Kar3E631APDB:1F9W | P43 | a=62.9Åc=153.6Å | 2.5Å | Molecularreplacement | |
| Ncd293-700PDB:1N6M | C2 | a=162.6Åb=66.6Åc=94.8Å | 2.5Å | Molecularreplacement | Rotationofstalkby75° |
| KIF1AAMPPNP:PDB:1VFVPDB:1VFWADP-AlFx:PDB:1VFXADP-Vi:PDB:1VFZ | AMPPNP1:P212121AMPPNP2:P212121ADP-AlFx:P212121ADP-Vi:P212121 | AMPPNP1:a=42.6Åb=55.2Åc=157.0ÅAMPPNP2:a=42.6Åb=55.5Åc=156.7ÅADP-AlFx:a=41.9Åb=54.6Åc=156.8ÅADP-Vi:a=41.5Åb=51.8Åc=157.0Å | AMPPNP1:1.85ÅAMPPNP2:2.2ÅADP-AlFx:2.6ÅADP-Vi:2.2Å | Molecularreplacement | MotorboundtoAMPPNP,ADP-AlFxorADP-Vi |
| KCBPPDB:1SDM | P21212 | a=95.7Åb=85.3Åc=44.5Å | 2.3Å | Molecularreplacement | |
| KinIPDB:1RY6 | P3221 | a=105.6Åc=84.8Å | 1.6Å | Molecularreplacement | Nonucleotideboundtomotor |
ContributedbyJ.Muller,J.KullandE.Mandelkow
IfyouusemethodsfromtheKinesinsite,weaskthatyoucitetheKinesinHomePageandauthors,ortheappropriatesourcepublicationinyourwork.
================ 蚂蚁淘在线 ================
免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容
版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。
非常满意 
