JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST000297/files/be1_fragment_201707.pdfhttps://storage.jpostdb.org/JPST000297/files/be1.rawprimaryOK200ProteomicsNobutaka HirokawaSus ScrofaMus Musculus (mouse)https://repository.jpostdb.org/entry/JPST000297Univ. of TokyojPOSTMicrotubules (MTs) are dynamic structures that are fundamental for cell morphogenesis and motility. MT-associated motors work efficiently to perform their functions. Unlike other motile kinesins, KIF2 catalytically depolymerizes MTs from the peeled protofilament end during ATP hydrolysis. However, the detailed mechanism by which KIF2 drives processive MT depolymerization remains unknown. To elucidate the catalytic mechanism, the transitional KIF2-tubulin complex during MT depolymerization was analyzed through multiple methods, including atomic force microscopy, size-exclusion chromatography, multi-angle light scattering, small-angle X-ray scattering, analytical ultracentrifugation, and mass spectrometry. The analyses outlined the conformation in which one KIF2core domain binds tightly to two tubulin dimers in the middle pre-hydrolysis state during ATP hydrolysis, a process critical for catalytic MT depolymerization. The X-ray crystallographic structure of the KIF2core domain displays the activated conformation that sustains the large KIF2-tubulin 1:2 complex.Mechanism of Catalytic Microtubule Depolymerization via KIF2-Tubulin Transitional Conformation.Ogawa Tadayuki T, Saijo Shinya S, Shimizu Nobutaka N, Jiang Xuguang X, Hirokawa Nobutaka Nmicrotubule catastrophe, tub, DTA1, beta-tubulin56D, alphat-1, alpha1tub, Dmbeta1, a1t, BETA 56D, epsilon-Tubulin, anon-EST:Liang-2.30, conformation, HK2, delta-Tubulin, anon-EST:Liang-1.59, Tubulin, betaTub1, Tubalpha1, alphatub, alphaT, alpha-tubulin, tubA84B, delta Tubulin, alpha-tub, DM1alpha, alpha-Tubulin, alpha1t, alpha-tubulin 84B, B1t, epsilon Tubulin, l(3)g3, KIF2, Kif2, l(3)84Bd, ms(3)nc33, BEST:LD32507, microtubule shortening, kif2-a, CDCBM3, alphaTUB, alphaTub, relational structural quality., alpha1, C530030B14Rik, Kns2, beta-tub, alpha Tubulin, betatub(56D), beta Tubulin, beta1tub, ALPHA 84C, CG9277, gamma-Tubulin, a-tub84B, beta[[1]]-tubulin, microtubule depolymerization during nuclear congression, DmelCG1913, beta[[1]] tubulin, alpha84B, Talpha1, beta1t, alphaTub1, T, beta-Tubulin, beta1, alpha1tub84B, aTub84B, beta56D, DmelCG9277, alphatub84B, beta, beta1-tub, DTB2, Tub, alpha-tub84B, Tuba84B, 1t, beta-Tub, alphaTub84, nc33, tubulin, beta1Tub, gamma Tubulin, M-kinesin, clone 1.59, alpha-Tub84B, tubalpha1, microtubule disassembly, clone 2.30, beta-Tub56D, alpha-Tub, CG1913, betaTub, beta1-Tubulintub, projections, alpha1tub, CPD photolyase activity, PP2A[[C]], PhrB photolyase activity, RAY, angular placement, Mbp1, Visible Light, betaTub1, pigmented epithelium, delta Tubulin, AU040256, alpha-Tubulin, alpha-tubulin 84B, DmelCG7109, Techniques, B1t, Light Microscopy, l(3)g3, PP2A 28D, Method, BEST:LD32507, hnu, PP2A[C], PP2, 3, CDCBM3, NUP96, DDON syndrome, Analysis, outer pigmented layer of retina, myd, epithelium, foton, deafness dystonia optic atrophy syndrome, Mass Spectrum Analysis, angular magnitude, Pp2A, beta1tub, ALPHA 84C, PP2a, Analyses, CG9277, Simple, DmelCG1913, force amplitude, hypoplasia, pigmented retina, alphaTub1, deafness-dystonia-optic neuronopathy (DDON) syndrome, T, beta-Tubulin, Mbp-1, MTS/PP2A, W, procedures, Hand-Held Microscopy, DNA cyclobutane dipyrimidine photolyase activity, SUPPRESSOR OF AUXIN RESISTANCE 3, alpha-tub84B, 1t, pigment epithelium of retina, Methodological Studies, beta1Tub, PP2A, Light-Chain Proteins, Hand-Held, papilla, pigmented retina epithelium, Pp2A-28D, dystonia, associated, alpha-Tub, 9530019H02Rik, ER2-6, DDP1, gamma, deafness-dystonia-optic neuronopathy syndrome, DTA1, PRE, alphat-1, PP2a 28D, a1t, BETA 56D, Radiation, anatomical protrusion, Kinesin Heavy-Chain, gyltl1b-b, deoxyribonucleic cyclobutane dipyrimidine photolyase activity, CG7109, lamina, flanges, Light, cellular morphogenesis, Procedure, pigmented retinal epithelium, alpha-tubulin, Optical Microscopy, Spectrum Analysis, Spectroscopy, LIGHT, epsilon Tubulin, KIF2, Kif2, retinal pigment, MDDGA6, mKIAA0609, shelf, DmPp2A-28D, dipyrimidine photolyase (photosensitive), beta1-Tubulin., deafness - dystonia - optic neuronopathy syndrome, END, Microtubule, plane angle, retinal pigment layer, C530030B14Rik, Visible Radiations, Kns2, fg, beta-tub, deafness dystonia optic neuronopathy syndrome, Visible Radiation, gyltl1b, Drives, HVEML, retinal pigmented epithelium, shelves, alpha84B, Talpha1, mdc1d, Spectrometry, expanded, alpha1tub84B, Methodological, beta56D, DmelCG9277, alphatub84B, Methodological Study, projection, ridge, Hand Held Microscopy, beta1-tub, DTB2, Tuba84B, beta-Tub, MDC1D, phr A photolyase activity, alphaTub84, light quantum, nc33, TIM8, DNA-photoreactivating enzyme, tubulin, enr, enlarged, spine, Microscopy, beta-Tub56D, progressive, CG1913, dystonia-deafness syndrome, betaTub, beta1-Tubulin, big, Procedures, conformation, anon-EST:Liang-2.30, lamellae, delta-Tubulin, Kinesin Heavy-Chain Protein, Kinesin Superfamily, anon-EST:Liang-1.59, dPP2A, Spectrum Analyses, photoreactivating enzyme activity, alphaT, tubA84B, process of organ, fractures, deoxyribodipyrimidine photolyase activity, froggy, stratum pigmentosum retinae, Gyltl1a, X-linked recessive, Ly113, protrusion, lamella, large, with blindness, reduced, H-ray, ms(3)nc33, Studies, Mass, kif2-a, Kinesin Light-Chain, stratum pigmentosa retinae, light, tiny, Technique, Mass Spectroscopy, alpha1, MOS3, PP2A-C, betatub(56D), deoxyribocyclobutadipyrimidine pyrimidine-lyase activity, MOHR-Tranebjaerg syndrome, Light-Chain Protein, beta Tubulin, PRECOCIOUS, amount of rotation, a-tub84B, deafness syndrome, Lichtquant, MDDGB6, beta[[1]] tubulin, beta1t, F23A5.3, ridges, dPP2A-C, Visible, angle, 5559, deafness dystonia optic neuronopathy syndrome (DDON), Study, Heavy-Chain Protein, BPFD#36, Optical, gamma Tubulin, and mental deficiency, M-kinesin, clone 1.59, great, tubalpha1, photon, TR2, clone 2.30, PP2Ac, Chromatographies, laminae, relational structural quality, HHT1, F23A5_3, small, beta-tubulin56D, Dmbeta1, epsilon-Tubulin, Mohr-Tranebjaerg syndrome, MTS, Mts, HK2, anatomical process, Proteins, Tubulin, Tubalpha1, alphatub, CD258, deafness-Dystonia-optic atrophy syndrome, alpha-tub, DM1alpha, alpha1t, stratum pigmentosum (retina), MODIFIER OF SNC1, l(3)84Bd, Simple Microscopy, Protein, deoxyribonucleic photolyase activity, PP2A C, techniques, DFN1, alphaTUB, alphaTub, Mass Spectrum Analyses, flange, organ process, Radiations, Mass Spectrum, l(2)02496, alpha Tubulin, RPE, RAB1C, deafness dystonia syndrome, underdeveloped, gamma-Tubulin, photolyase activity, Compound Microscopy, HVEM-L, beta[[1]]-tubulin, DDP, Photoradiation, l(2)s5286, beta1, aTub84B, retinal pigment epithelium, Kinesin, LTg, beta, p. pigmentosa retinae, Tub, processes, process, Superfamily, Photoradiations, Compound, Kinesin Light-Chain Protein, alpha-Tub84B, processus, deoxyribonucleate pyrimidine dimer lyase (photosensitive), ORW1, Kinesin Light-Chain Proteins, dystonia deafness syndrome, methodologyCT139, X83316, mcak, MCAK, 4930402F02Rik, KNSL6., ESTM5, Knsl6C530030B14Rik, Kns2, kif2-a, CDCBM3, KIF2, Kif2, M-kinesin, HK2.falseX-link of KIF2C-tubulin complex with EDCKIF2-tubulin complex was crosslinked with EDC, trypsinized, and analyzed by LC-MS.Wed Sep 13 00:00:00 BST 2017PXD00715210090982328903043