A "slow" homotetrameric kinesin-related motor protein purified from Drosophila embryos.
A "slow" homotetrameric kinesin-related motor protein purified from Drosophila embryos.
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发表时间：
1994-09
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通讯作者：
Douglas G. Cole;W. Saxton;Kathy B. Sheehan;J. Scholey
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作者：
Douglas G. Cole;W. Saxton;Kathy B. Sheehan;J. Scholey
泛驱动蛋白肽抗体(Cole， D. G., Cande， W. Z., Baskin， R. J, Skoufias， D. A., Hogan， C. J, and Scholey， J. M.(1992) .细胞科学101,291-301；Sawin， K. E., Mitchinson， T. J., Wordeman， L. G. (1992) J.细胞科学，101(03)-313)，从果蝇胚胎细胞质中鉴定和分离运动蛋白相关蛋白（KRPs）。这些KRPs与经AMP-PNP（腺苷-5′-酰基酰亚胺二磷酸）处理的细胞质溶胶聚合的微管（MTs）共同沉积，其中一个KRP130从胚胎MTs的ATP洗脱液中进一步纯化。纯化后的KRP130表现为由四个130-kDa多肽亚基组成的同四聚体复合物，表现出“缓慢”的正端定向运动活性，能够以0.04 +/- 0.01微米/秒的速度移动单个MTs。KRP130的130-kDa亚基与单克隆和多克隆抗体的反应性进行了测试，这些抗体是针对各种激酶蛋白超家族成员的特异性抗体。结果表明，KRP130亚基与Xenopus Eg5 （Sawin， K. E., Le Guellec， K. L., Philippe， M., Mitchinson， T. J. (1992) Nature 359, 540-543）有亲缘关系。因此，KRP130似乎是第一个从原生组织中作为多聚体运动复合物纯化的果蝇KRP，也是任何生物体中BimC亚家族的第一个成员。
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.