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Purification, crystallization and preliminary X-ray diffraction analysis of the catalytic domain of adenylyl cyclase Rv1625c from Mycobacterium tuberculosis

Ketkar, Amit D and Shenoy, Avinash R and Kesavulu, Muppuru M and Visweswariah, Sandhya S and Suguna, Kaza (2004) Purification, crystallization and preliminary X-ray diffraction analysis of the catalytic domain of adenylyl cyclase Rv1625c from Mycobacterium tuberculosis. In: Acta Crystallographica Section D: Biological Crystallography, 60 (Part 2). pp. 371-373.

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Abstract

The Rv1625c gene product is an adenylyl cyclase identiÆed 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 \rightarrow 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 \AA . X-ray diffraction data were collected to a resolution of 3.4 \AA and the structure has been solved by the molecular-replacement method using a previously built theoretical model of the protein as the search molecule.

Item Type: Journal Article
Additional Information: The copyright belongs to International Union of Crystallography.
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Division of Biological Sciences > Molecular Reproduction, Development & Genetics (formed by the merger of DBGL and CRBME)
Date Deposited: 30 May 2006
Last Modified: 19 Sep 2010 04:27
URI: http://eprints.iisc.ernet.in/id/eprint/7109

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