酵母Sec2p（一种酵母Rab GTP酶Sec4p的鸟嘌呤核苷酸交换因子）的结晶及晶体学分析。
Crystallization and crystallographic analysis of yeast Sec2p, a guanine nucleotide-exchange factor for the yeast Rab GTPase Sec4p.
作者信息
Sato Yusuke, Fukai Shuya, Nureki Osamu
机构信息
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama-shi, Kanagawa 226-8501, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Jul 1;63(Pt 7):616-9. doi: 10.1107/S1744309107027121. Epub 2007 Jun 22.
Sec2p is a guanine nucleotide-exchange factor (GEF) for the yeast Rab GTPase Sec4p. Sec2p accelerates GDP release from Sec4p and promotes GDP-GTP exchange for Sec4p activation. In order to elucidate this nucleotide-exchange mechanism using X-ray crystallography, three constructs of native Sec2p (Sec2(1-160)p, Sec2(18-160)p and Sec2(31-160)p) and three constructs of selenomethionine-labelled Sec2p [Sec2(31-160)p, Sec2(31-160)p (M115L) and Sec2(31-160)p (M115L, K121M, T142M)] were crystallized. These six crystals diffracted to 8.8, 4.8, 2.6, 4.0, 3.3 and 3.0 A resolution, respectively. The data set from the SeMet-labelled Sec2(31-160)p (M115L, K121M, T142M) crystal was processed for SAD phasing; the crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 101.9, b = 176.6, c = 181.5 A.
Sec2p是酵母Rab GTP酶Sec4p的鸟嘌呤核苷酸交换因子（GEF）。Sec2p加速Sec4p中GDP的释放，并促进GDP与GTP的交换以激活Sec4p。为了利用X射线晶体学阐明这种核苷酸交换机制，制备了天然Sec2p的三种构建体（Sec2(1-160)p、Sec2(18-160)p和Sec2(31-160)p）以及硒代甲硫氨酸标记的Sec2p的三种构建体[Sec2(31-160)p、Sec2(31-160)p (M115L)和Sec2(31-160)p (M115L, K121M, T142M)]并使其结晶。这六块晶体的衍射分辨率分别为8.8、4.8、2.6、4.0、3.3和3.0埃。对来自硒代甲硫氨酸标记的Sec2(31-160)p (M115L, K121M, T142M)晶体的数据集进行单波长反常散射（SAD）相位分析；该晶体属于空间群P2(1)2(1)2(1)，晶胞参数a = 101.9，b = 176.6，c = 181.5埃。
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