Expression, purification and spectroscopic characterization of the cytochrome P450CYP121 from Mycobacterium tuberculosis

McLean, Kirsty J., Cheesman, Myles R., Rivers, Stuart L., Richmond, Alison, Leys, David, Chapman, Stephen K., Reid, Graeme A., Price, Nicholas C., Kelly, Sharon M., Clarkson, John, Smith, W. Ewen and Munro, Andrew W. (2002) Expression, purification and spectroscopic characterization of the cytochrome P450CYP121 from Mycobacterium tuberculosis. Journal of Inorganic Biochemistry, 91 (4). pp. 527-541. ISSN 0162-0134

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Abstract

The CYP121 gene from the pathogenic bacterium Mycobacterium tuberculosis has been cloned and expressed in Escherichia coli, and the protein purified to homogeneity by ion exchange and hydrophobic interaction chromatography. The CYP121 gene encodes a cytochrome P450 enzyme (CYP121) that displays typical electronic absorption features for a member of this superfamily of hemoproteins (major Soret absorption band at 416.5 nm with alpha and beta bands at 565 and 538 nm, respectively, in the oxidized form) and which binds carbon monoxide to give the characteristic Soret band shift to 448 run. Resonance Raman, EPR and MCD spectra show the protein to be predominantly low-spin and to have a typical cysteinate- and water-ligated b-type heme iron. CD spectra in the far UV region describe a mainly alpha helical conformation, but the visible CD spectrum shows a band of positive sign in the Soret region, distinct from spectra for other P450s recognized thus far. CYP121 binds very tightly to a range of azole antifungal drugs (e.g. clotrimazole, miconazole), suggesting that it may represent a novel target for these antibiotics in the M. tuberculosis pathogen. (C) 2002 Elsevier Science Inc. All rights reserved.

Item Type:	Article
Additional Information:	12th International Conference on Advances in the Inorganic Biochemistry of Cytochrome P450 SEP, 2001 LA GRANDE MOTTE, FRANCE
Uncontrolled Keywords:	sdg 3 - good health and well-being ,/dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being
Faculty \ School:	Faculty of Science > School of Chemistry (former - to 2024)
UEA Research Groups:	Faculty of Science > Research Groups > Biophysical Chemistry (former - to 2017) Faculty of Science > Research Groups > Chemistry of Life Processes Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry Faculty of Science > Research Groups > Chemistry of Light and Energy
Depositing User:	Rachel Smith
Date Deposited:	10 May 2011 11:09
Last Modified:	24 Sep 2024 10:18
URI:	https://ueaeprints.uea.ac.uk/id/eprint/30086
DOI:	10.1016/S0162-0134(02)00479-8

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