M B Fisher1, Y M Zheng, A E Rettie. 1. Department of Medicinal Chemistry, University of Washington, Seattle, Washington, 98195, USA.
Abstract
Rabbit CYP4B1 was incubated with a series of fatty acid and hydrocarbon substrates and metabolites were identified by gas chromatography and gas chromatography/mass spectrometry. C-7 to C-10 n-alkyl fatty acids were preferentially hydroxylated at the terminal carbon (omega/omega-1 = 1.1-7.4) with turnover numbers of 1-11 min-1. The C-7 to C-10 n-alkyl hydrocarbons exhibited turnover numbers of 11-33 min-1 for the corresponding reactions and even higher regioselectivities for hydroxylation at the thermodynamically disfavored site (omega/omega-1 = 1.6-23). These results demonstrate a functional link between CYP4B1 and other CYP4 fatty acid hydroxylases, and show further that CYP4B1's unusual positional specificity is not dictated by the presence of a carboxylate (or polar) anchor on the substrate. This suggests the presence of a dominant hydrocarbon binding site which effectively restricts the access of short-medium chain n-alkyl substrates to the perferryl species in the active site of rabbit CYP4B1. Copyright 1998 Academic Press.
Rabbitn class="Gene">CYP4B1 was incubated with a series of fatty acid and hydrocarbon substrates and metabolites were identified by gas chromatography and gas chromatography/mass spectrometry. C-7 to C-10 n-alkyl fatty acids were preferentially hydroxylated at the terminal carbon (omega/omega-1 = 1.1-7.4) with turnover numbers of 1-11 min-1. The C-7 to C-10 n-alkyl hydrocarbons exhibited turnover numbers of 11-33 min-1 for the corresponding reactions and even higher regioselectivities for hydroxylation at the thermodynamically disfavored site (omega/omega-1 = 1.6-23). These results demonstrate a functional link between CYP4B1 and other CYP4 fatty acid hydroxylases, and show further that CYP4B1's unusual positional specificity is not dictated by the presence of a carboxylate (or polar) anchor on the substrate. This suggests the presence of a dominant hydrocarbon binding site which effectively restricts the access of short-medium chain n-alkyl substrates to the perferryl species in the active site of rabbitCYP4B1. Copyright 1998 Academic Press.
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