Open AccessCloning, overexpression, purification, crystallization and preliminary X‐ray diffraction analysis of an inositol monophosphatase family protein (SAS2203) from Staphylococcus aureus MSSA476
Author(s) -
Bhattacharyya Sudipta,
Dutta Debajyoti,
Ghosh Ananta Kumar,
Das Amit Kumar
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111003496
Subject(s) - crystallization , cloning (programming) , staphylococcus aureus , inositol , chemistry , microbiology and biotechnology , diffraction , crystallography , biology , biochemistry , genetics , bacteria , physics , optics , receptor , organic chemistry , computer science , programming language
The gene product of the sas 2203 ORF of Staphylococcus aureus MSSA476 encodes a 30 kDa molecular‐weight protein with a high sequence resemblance (29% identity) to tetrameric inositol monophosphatase from Thermotoga maritima . The protein was cloned, expressed, purified to homogeneity and crystallized. Crystals appeared in several conditions and good diffraction‐quality crystals were obtained from 0.2 M Li 2 SO 4 , 20% PEG 3350, 0.1 M HEPES pH 7.0 using the sitting‐drop vapour‐diffusion method. A complete diffraction data set was collected to 2.6 Å resolution using a Rigaku MicroMax‐007 HF Cu K α X‐ray generator and a Rigaku R‐AXIS IV ++ detector. The diffraction data were consistent with the orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 49.98, b = 68.35, c = 143.79 Å, α = β = γ = 90°, and the crystal contained two molecules in the asymmetric unit.