Open AccessExpression, purification and crystallization of acetyl‐CoA hydrolase from Neisseria meningitidis
Author(s) -
Khandokar Yogesh B.,
Londhe Avinash,
Patil Shilpa,
Forwood Jade K.
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113028042
Subject(s) - neisseria meningitidis , streptococcus pneumoniae , hydrolase , microbiology and biotechnology , crystallization , bacteria , enzyme , biology , biochemistry , escherichia coli , chemistry , gene , antibiotics , genetics , organic chemistry
Neisseria meningitidis is the causative microorganism of many human diseases, including bacterial meningitis; together with Streptococcus pneumoniae , it accounts for approximately 80% of bacterial meningitis infections. The emergence of antibiotic‐resistant strains of N. meningitidis has created a strong urgency for the development of new therapeutics, and the high‐resolution structural elucidation of enzymes involved in cell metabolism represents a platform for drug development. Acetyl‐CoA hydrolase is involved in multiple functions in the bacterial cell, including membrane synthesis, fatty‐acid and lipid metabolism, gene regulation and signal transduction. Here, the first recombinant protein expression, purification and crystallization of a hexameric acetyl‐CoA hydrolase from N. meningitidis are reported. This protein was crystallized using the hanging‐drop vapour‐diffusion technique at pH 8.5 and 290 K using ammonium phosphate as a precipitant. Optimized crystals diffracted to 2.0 Å resolution at the Australian Synchrotron and belonged to space group P 2 1 3 (unit‐cell parameters a = b = c = 152.2 Å), with four molecules in the asymmetric unit.