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Human cytochrome P450 4F11: heterologous expression in bacteria, purification, and characterization of catalytic function.


ABSTRACT: Human cytochrome P450 (P450) 4F11 is still considered an "orphan" because its function is not well characterized. A bacterial expression system was developed for human P450 4F11, producing approximately 230nmol P450 from a 3-l culture of Escherichia coli. P450 4F11 was purified and utilized for untargeted substrate searches in human liver extract using a liquid chromatography/mass spectrometry-based metabolomic and isotopic labeling approach (Tang et al., 2009 [19]). Four fatty acids-palmitic, oleic, arachidonic, and docosahexaenoic-were identified in human liver and verified as substrates of P450 4F11. The products were characterized as omega-hydroxylated fatty acids by gas chromatography-mass spectrometry analysis of their trimethylsilyl derivatives. Kinetic analysis of the oxidation products confirmed that the fatty acids are substrates oxidized by P450 4F11. P450 4F11 also exhibited low activity for some drug N-demethylation reactions but none for activation of several pro-carcinogens.

SUBMITTER: Tang Z 

PROVIDER: S-EPMC2812615 | biostudies-literature | 2010 Feb

REPOSITORIES: biostudies-literature

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Human cytochrome P450 4F11: heterologous expression in bacteria, purification, and characterization of catalytic function.

Tang Zhongmei Z   Salamanca-Pinzón Sandra Giovanna SG   Wu Zhong-Liu ZL   Xiao Yi Y   Guengerich F Peter FP  

Archives of biochemistry and biophysics 20091120 1


Human cytochrome P450 (P450) 4F11 is still considered an "orphan" because its function is not well characterized. A bacterial expression system was developed for human P450 4F11, producing approximately 230nmol P450 from a 3-l culture of Escherichia coli. P450 4F11 was purified and utilized for untargeted substrate searches in human liver extract using a liquid chromatography/mass spectrometry-based metabolomic and isotopic labeling approach (Tang et al., 2009 [19]). Four fatty acids-palmitic, o  ...[more]

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