title: Purification, crystallization and preliminary X-ray diffraction analysis of the catalytic domain of adenylyl cyclase Rv1625c from Mycobacterium tuberculosis.
abstract: The Rv1625c gene product is an adenylyl cyclase identified in the genome of Mycobacterium tuberculosis strain H37Rv. It shows sequence similarity to the mammalian nucleotide cyclases and functions as a homodimer, with two substrate-binding sites at the dimer interface. A mutant form of the catalytic domain of this enzyme, K296E/F363R/D365C (KFD-->ERC), was overexpressed in Escherichia coli cells in a soluble form. Crystals were obtained using the hanging-drop vapour-diffusion method with PEG 8000 as a precipitant. The protein crystallized in space group P4(1), with unit-cell parameters a = b = 71.25, c = 44.51 A. X-ray diffraction data were collected to a resolution of 3.4 A and the structure has been solved by the molecular-replacement method using a previously built theoretical model of the protein as the search molecule.
authors.affiliation: Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore 560012, India.
authors.forename: Amit D
authors.initials: AD
authors.lastname: Ketkar
authors.forename: Avinash R
authors.initials: AR
authors.lastname: Shenoy
authors.forename: Muppuru M
authors.initials: MM
authors.lastname: Kesavulu
authors.forename: Sandhya S
authors.initials: SS
authors.lastname: Visweswariah
authors.forename: Kaza
authors.initials: K
authors.lastname: Suguna
mesh_terms: D000262:Adenylyl Cyclases / Q000737:chemistry*; D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D003460:Crystallization; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D011092:Polyethylene Glycols / Q000737:chemistry; D017434:Protein Structure, Tertiary; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D011092:Polyethylene Glycols; D000262:Adenylyl Cyclases