Open AccessCrystallization and crystallographic analysis of yeast Sec2p, a guanine nucleotide‐exchange factor for the yeast Rab GTPase Sec4p
Author(s) -
Sato Yusuke,
Fukai Shuya,
Nureki Osamu
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107027121
Subject(s) - guanine nucleotide exchange factor , gtpase , rab , gtp' , yeast , crystallography , nucleotide , crystallization , guanine , chemistry , stereochemistry , biochemistry , gene , organic chemistry , enzyme
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 Å 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 P 2 1 2 1 2 1 , with unit‐cell parameters a = 101.9, b = 176.6, c = 181.5 Å.