Cole D G, Saxton W M, Sheehan K B, Scholey J M
Section of Molecular and Cellular Biology, University of California, Davis 95616.
J Biol Chem. 1994 Sep 16;269(37):22913-6.
Pan-kinesin peptide antibodies (Cole, D. G., Cande, W. Z., Baskin, R. J., Skoufias, D. A., Hogan, C. J., and Scholey, J. M. (1992) J. Cell Sci. 101, 291-301; Sawin, K. E., Mitchinson, T. J., and Wordeman, L. G. (1992) J. Cell Sci. 101, 303-313) were used to identify and isolate kinesin-related proteins (KRPs) from Drosophila melanogaster embryonic cytosol. These KRPs cosedimented with microtubules (MTs) polymerized from cytosol treated with AMP-PNP (adenyl-5'-yl imidodiphosphate), and one of them, KRP130, was further purified from ATP eluates of the embryonic MTs. Purified KRP130 behaves as a homotetrameric complex composed of four 130-kDa polypeptide subunits which displays a "slow" plus-end directed motor activity capable of moving single MTs at 0.04 +/- 0.01 microns/s. The 130-kDa subunit of KRP130 was tested for reactivity with monoclonal and polyclonal antibodies that are specific for various members of the kinesin superfamily. Results indicate that the KRP130 subunit is related to Xenopus Eg5 (Sawin, K. E., Le Guellec, K. L., Philippe, M., Mitchinson, T. J. (1992) Nature 359, 540-543), a member of the BimC subfamily of kinesins. Therefore, KRP130 appears to be the first Drosophila KRP, and the first member of the BimC subfamily in any organism, to be purified from native tissue as a multimeric motor complex.
泛驱动蛋白肽抗体（科尔，D.G.，坎德，W.Z.，巴斯金，R.J.，斯科菲亚斯，D.A.，霍根，C.J.，以及肖利，J.M.（1992年）《细胞科学杂志》101卷，291 - 301页；索温，K.E.，米奇森，T.J.，以及沃德曼，L.G.（1992年）《细胞科学杂志》101卷，303 - 313页）被用于从黑腹果蝇胚胎胞质溶胶中鉴定和分离驱动蛋白相关蛋白（KRPs）。这些KRPs与用AMP - PNP（腺苷 - 5'-亚氨基二磷酸）处理的胞质溶胶聚合形成的微管（MTs）共同沉降，其中一个名为KRP130的蛋白，从胚胎MTs的ATP洗脱液中进一步纯化得到。纯化后的KRP130表现为一种同四聚体复合物，由四个130 kDa的多肽亚基组成，具有“缓慢”的向正极定向的运动活性，能够以0.04±0.01微米/秒的速度移动单个MTs。对KRP130的130 kDa亚基与针对驱动蛋白超家族各成员的单克隆和多克隆抗体的反应性进行了测试。结果表明，KRP130亚基与非洲爪蟾Eg5（索温，K.E.，勒盖莱克，K.L.，菲利普，M.，米奇森，T.J.（1992年）《自然》359卷，540 - 543页）相关，Eg5是驱动蛋白BimC亚家族的成员。因此，KRP130似乎是第一个从黑腹果蝇中分离得到的KRP，也是在任何生物体中第一个作为多聚体运动复合物从天然组织中纯化得到的BimC亚家族成员。
