title: Crystallization and preliminary X-ray crystallographic analysis of the oxysterol-binding protein Osh3 from Saccharomyces cerevisiae.
abstract: Oxysterol-binding protein (OSBP) related proteins (ORPs) are conserved from yeast to humans and are implicated in regulation of sterol homeostasis and in signal transduction pathways. Osh3 of Saccharomyces cerevisiae is a pleckstrin-homology (PH) domain-containing ORP member that regulates phosphoinositide metabolism at endoplasmic reticulum-plasma membrane contact sites. The N-terminal PH domain of Osh3 was purified and crystallized as a lysozyme fusion and the resulting crystal diffracted to 2.3 Å resolution. The crystal belonged to the monoclinic space group C2, with unit-cell parameters a=98.03, b=91.31, c=84.13 Å, β=81.41°. With two molecules in the asymmetric unit, the Matthews coefficient was 3.13 Å3 Da(-1). Initial attempts to solve the structure by molecular-replacement techniques using T4 lysozyme as a search model were successful. The C-terminal OSBP-related domain (OBD) of Osh3 was crystallized by the vapour-diffusion method and the resulting crystal diffracted to 1.5 Å resolution. The crystal was orthorhombic, belonging to space group P2(1)2(1)2(1), with unit-cell parameters a=41.57, b=87.52, c=100.58 Å. With one molecule in the asymmetric unit, the Matthews coefficient was 2.01 Å3 Da(-1). Initial attempts to solve the structure by the single-wavelength anomalous dispersion technique using bromine were successful.
authors.affiliation: College of Pharmacy, Chonnam National University, Gwangju 500-757, Republic of Korea.
authors.forename: Junsen
authors.initials: J
authors.lastname: Tong
authors.forename: Huiseon
authors.initials: H
authors.lastname: Yang
authors.forename: Subin
authors.initials: S
authors.lastname: Ha
authors.forename: Youngjin
authors.initials: Y
authors.lastname: Lee
authors.forename: Soo Hyun
authors.initials: SH
authors.lastname: Eom
authors.forename: Young Jun
authors.initials: YJ
authors.lastname: Im
mesh_terms: D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; C432936:OSH3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins