牛 Rho 激酶 Rho 结合结构域的结晶及初步晶体学分析

Crystallization and preliminary crystallographic analysis of the Rho-binding domain of bovine Rho-kinase.

作者信息

Ihara K, Shimizu T, Maesaki R, Okada K, Amano M, Kaibuchi K, Hakoshima T

机构信息

Division of Structural Biology, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan.

出版信息

Acta Crystallogr D Biol Crystallogr. 2000 Aug;56(Pt 8):1042-4. doi: 10.1107/s0907444900007174.

DOI:10.1107/s0907444900007174

PMID:10944348

Abstract

Rho-kinase binds to a small GTPase Rho in a GTP-dependent manner and regulates many cytoskeletal events in the cell. The minimum region of bovine Rho-kinase sufficient for Rho-binding was expressed as a fusion protein with glutathione S-transferase. After removal of the glutathione S-transferase, thin plate crystals were obtained. The selenomethionine-substituted protein was introduced and crystallized, as was the native protein. The crystals of the Rho-binding domain of Rho-kinase belong to the space group C2, with unit-cell parameters a = 148.0 (2), b = 26.1 (1), c = 39.6 (1) A, beta = 90.3 (1) degrees. The crystals diffract to a resolution beyond 1.5 A.

摘要

Rho激酶以GTP依赖的方式与小GTP酶Rho结合，并调节细胞内许多细胞骨架事件。将牛Rho激酶中足以与Rho结合的最小区域表达为与谷胱甘肽S-转移酶的融合蛋白。去除谷胱甘肽S-转移酶后，获得了薄板晶体。引入了硒代甲硫氨酸取代的蛋白质并使其结晶，天然蛋白质也是如此。Rho激酶的Rho结合结构域的晶体属于空间群C2，晶胞参数为a = 148.0（2），b = 26.1（1），c = 39.6（1）Å，β = 90.3（1）度。这些晶体的衍射分辨率超过1.5 Å。