Crystallization and crystallographic analysis of yeast Sec2p, a guanine nucleotide-exchange factor for the yeast Rab GTPase Sec4p.
Crystallization and crystallographic analysis of yeast Sec2p, a guanine nucleotide-exchange factor for the yeast Rab GTPase Sec4p.
复制标题
DOI：
10.1107/s1744309107027121
复制
发表时间：
2007-07
期刊：
影响因子：
--
通讯作者：
Yusuke Sato;S. Fukai;O. Nureki
中科院分区：
文献类型：
--
作者：
Yusuke Sato;S. Fukai;O. Nureki
Sec 2 p是一种鸟嘌呤核苷酸交换因子（GEF），用于酵母菌Rab GTc 4p。Sec 2 p加速Sec 4p释放GDP，并促进Sec 4p激活的GDP-GTP交换。为了使用X射线晶体学阐明这种核苷酸交换机制，使用了天然Sec 2 p的三种构建体。（Sec 2（1-160）p、Sec 2（18-160）p和Sec 2（31-160）p）和硒代甲硫氨酸标记的Sec 2 p的三种构建体[Sec 2（31 - 160）p、Sec 2（31-160）p（M115 L）和Sec 2（31-160）p（M115 L，K121 M，T142 M）]的结晶。这六个晶体分别衍射到8.8、4.8、2.6、4.0、3.3和3.0 A分辨率。对来自SeMet标记的Sec 2（31-160）p（M115 L，K121 M，T142 M）晶体的数据集进行SAD定相处理;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 A。
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.