title: Expression, purification and crystallization of acetyl-CoA hydrolase from Neisseria meningitidis.
abstract: 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 P2(1)3 (unit-cell parameters a = b = c = 152.2 Å), with four molecules in the asymmetric unit.
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Boorooma Street, Wagga Wagga, New South Wales 2650, Australia.
authors.forename: Yogesh B
authors.initials: YB
authors.lastname: Khandokar
authors.forename: Avinash
authors.initials: A
authors.lastname: Londhe
authors.forename: Shilpa
authors.initials: S
authors.lastname: Patil
authors.forename: Jade K
authors.initials: JK
authors.lastname: Forwood
mesh_terms: D000104:Acetyl-CoA Hydrolase / Q000737:chemistry* / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D009345:Neisseria meningitidis / Q000201:enzymology*; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D000104:Acetyl-CoA Hydrolase
keywords: Neisseria meningitidis; acetyl-CoA hydrolase; hotdog fold; thioesterase