title: Expression, purification, crystallization and preliminary X-ray analysis of the acyl carrier protein synthase (acpS) from Mycobacterium tuberculosis.
abstract: Acyl carrier protein synthase (acpS) catalyzes the formation of holo-ACP, which mediates the transfer of acyl fatty-acid intermediates during the biosynthesis of fatty acids and lipids. An expression and purification system for the Mycobacterium tuberculosis (Mtb) acpS has been established that yields approximately 15 mg l(-1) of the enzyme in soluble form. The purified enzyme has been crystallized by the vapour-diffusion method using 2-propanol as a precipitant. The original crystal size has been improved significantly by the addition of glycerol to the mother liquor. Mtb acpS crystals belong to the space group R3, with unit-cell parameters a = b = 68.53, c = 85.9 A. Native data have been collected under cryogenic conditions; phase resolution by molecular replacement and selenomethionine-aided multi-wavelength anomalous dispersion techniques is ongoing.
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Sidharth
authors.initials: S
authors.lastname: Chopra
authors.forename: Saurabh Kumar
authors.initials: SK
authors.lastname: Singh
authors.forename: Sushil Prasad
authors.initials: SP
authors.lastname: Sati
authors.forename: Anand
authors.initials: A
authors.lastname: Ranganathan
authors.forename: Amit
authors.initials: A
authors.lastname: Sharma
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D011487:Protein Conformation; D017855:Transferases (Other Substituted Phosphate Groups) / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D017855:Transferases (Other Substituted Phosphate Groups); C021364:holo-(acyl-carrier-protein) synthase