Characterization of active site structure in cyp121 a cytochrome p450 essential for viability of mycobacterium tuberculosis h37rv

Smith, W.E. and Lewis, D.G. and Dunford, A.J. and Seward, H.E. and Neeli, R. and Cheesman, M.R. and Marsollier, L. (2008) Characterization of active site structure in cyp121 a cytochrome p450 essential for viability of mycobacterium tuberculosis h37rv. Journal of Biological Chemistry, 283 (48). pp. 33406-33416. ISSN 1083-351X (http://dx.doi.org/10.1074/jbc.M802115200)

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Abstract

Mycobacterium tuberculosis (Mtb) cytochrome P450 gene CYP121 is shown to be essential for viability of the bacterium in vitro by gene knock-out with complementation. Production of CYP121 protein in Mtb cells is demonstrated. Minimum inhibitory concentration values for azole drugs against Mtb H37Rv were determined, the rank order of which correlated well with Kd values for their binding to CYP121. Solution-state spectroscopic, kinetic, and thermodynamic studies and crystal structure determination for a series of CYP121 active site mutants provide further insights into structure and biophysical features of the enzyme. Pro346 was shown to control heme cofactor conformation, whereas Arg386 is a critical determinant of heme potential, with an unprecedented 280-mV increase in heme iron redox potential in a R386L mutant. A homologous Mtb redox partner system was reconstituted and transported electrons faster to CYP121 R386L than to wild type CYP121. Heme potential was not perturbed in a F338H mutant, suggesting that a proposed P450 superfamily-wide role for the phylogenetically conserved phenylalanine in heme thermodynamic regulation is unlikely. Collectively, data point to an important cellular role for CYP121 and highlight its potential as a novel Mtb drug target.

  • Item type:Article
    ID code:19413
    Dates:
    Date
    Event
    28 November 2008
    Published
    Notes:Strathprints' policy is to record up to 8 authors per publication, plus any additional authors based at the University of Strathclyde. More authors may be listed on the official publication than appear in the Strathprints' record.
    Subjects:Science > Chemistry
    Department:Faculty of Science > Pure and Applied Chemistry
    Depositing user: Strathprints Administrator
    Date deposited:19 May 2010 13:20
    Last modified:10 Apr 2024 15:53
    URI:https://strathprints.strath.ac.uk/id/eprint/19413
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