Open AccessPurification, crystallization and preliminary X‐ray analysis of 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase of Streptococcus pneumoniae
Author(s) -
Zhang Liping,
Feng Lingling,
Zhou Li,
Gui Jie,
Wan Jian,
Hu Xiaopeng
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110036481
Subject(s) - reductase , orthorhombic crystal system , streptococcus pneumoniae , crystallization , coenzyme a , solvent , crystallography , chemistry , crystal (programming language) , biochemistry , crystal structure , enzyme , antibiotics , organic chemistry , programming language , computer science
Class II 3‐hydroxy‐3‐methylglutaryl‐coenzyme A (HMG‐CoA) reductases are potential targets for novel antibiotic development. In order to obtain a precise structural model for use in virtual screening and inhibitor design, HMG‐CoA reductase of Streptococcus pneumoniae was cloned, overexpressed and purified to homogeneity using Ni–NTA affinity chromatography. Crystals were obtained using the hanging‐drop vapour‐diffusion method. A complete data set was collected from a single frozen crystal on a home X‐ray source. The crystal diffracted to 2.3 Å resolution and belonged to the orthorhombic space group C 222 1 , with unit‐cell parameters a = 773.4836, b = 90.3055, c = 160.5592 Å, α = β = γ = 90°. Assuming the presence of two molecules in the asymmetric unit, the solvent content was estimated to be 54.1% ( V M = 2.68 Å 3 Da −1 ).