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13C-Methyl isocyanide as an NMR probe for cytochrome P450 active sites.


ABSTRACT: The cytochromes P450 (CYPs) play a central role in many biologically important oxidation reactions, including the metabolism of drugs and other xenobiotic compounds. Because they are often assayed as both drug targets and anti-targets, any tools that provide: (a) confirmation of active site binding and (b) structural data, would be of great utility, especially if data could be obtained in reasonably high throughput. To this end, we have developed an analog of the promiscuous heme ligand, cyanide, with a (13)CH(3)-reporter attached. This (13)C-methyl isocyanide ligand binds to bacterial (P450cam) and membrane-bound mammalian (CYP2B4) CYPs. It can be used in a rapid 1D experiment to identify binders, and provides a qualitative measure of structural changes in the active site.

SUBMITTER: McCullough CR 

PROVIDER: S-EPMC3870883 | biostudies-literature | 2009 Mar

REPOSITORIES: biostudies-literature

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13C-Methyl isocyanide as an NMR probe for cytochrome P450 active sites.

McCullough Christopher R CR   Pullela Phani Kumar PK   Im Sang-Choul SC   Waskell Lucy L   Sem Daniel S DS  

Journal of biomolecular NMR 20090206 3


The cytochromes P450 (CYPs) play a central role in many biologically important oxidation reactions, including the metabolism of drugs and other xenobiotic compounds. Because they are often assayed as both drug targets and anti-targets, any tools that provide: (a) confirmation of active site binding and (b) structural data, would be of great utility, especially if data could be obtained in reasonably high throughput. To this end, we have developed an analog of the promiscuous heme ligand, cyanide  ...[more]

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