Expression and characterization of cytochrome P4504A7 in E.coli

P. A. Loughran, L. J. Roman, B. S S Masters

Research output: Contribution to journalArticle

Abstract

The cytochromes P450 (P450) constitute a family of hemeproteins that serve as monooxygenases for exogenous and endogenous substrates. The P4504A subfamily, consisting of eicosanoid and fatty acid hydroxylases, plays a role in the metabolism of arachidonic acid in lung, liver and kidney. This metabolism of arachidonic acid produces 20-hydroxyeicosatetraenoic acid (20-HETE), shown to be a potent vasoconstrictor and, consequently, could be involved in hemodynamic regulation (Sawamura et al., 1993). The P4504A family members from rabbit, 4A4, 4A5, 4A6, and 4A7, are 85% homologous in amino acid sequence but have been shown to exhibit differing substrate specificities (Roman et al., 1993). To determine the structural and functional characteristics of the P4504A7 substrate binding/recognition site, production of large amounts of enzyme in an E.coli expression was accomplished (20nmoles/liter) as a chimera with the first ten amino acids of bovine P45017a. Kinetic characterization determined that the primary substrate, lauric acid, was metabolized at a turnover number of 20min1, with a Km of 1.6nM. Characterization of other substrates will be presented.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - 1997

Fingerprint

lauric acid
Cytochromes
cytochromes
Mixed Function Oxygenases
arachidonic acid
Arachidonic Acid
Escherichia coli
Hemeproteins
vasoconstrictor agents
Amino Acid Sequence Homology
dodecanoic acid
metabolism
eicosanoids
Eicosanoids
chimerism
Vasoconstrictor Agents
Substrates
hemodynamics
substrate specificity
Substrate Specificity

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Loughran, P. A., Roman, L. J., & Masters, B. S. S. (1997). Expression and characterization of cytochrome P4504A7 in E.coli. FASEB Journal, 11(9).

Expression and characterization of cytochrome P4504A7 in E.coli. / Loughran, P. A.; Roman, L. J.; Masters, B. S S.

In: FASEB Journal, Vol. 11, No. 9, 1997.

Research output: Contribution to journalArticle

Loughran, PA, Roman, LJ & Masters, BSS 1997, 'Expression and characterization of cytochrome P4504A7 in E.coli', FASEB Journal, vol. 11, no. 9.
Loughran PA, Roman LJ, Masters BSS. Expression and characterization of cytochrome P4504A7 in E.coli. FASEB Journal. 1997;11(9).
Loughran, P. A. ; Roman, L. J. ; Masters, B. S S. / Expression and characterization of cytochrome P4504A7 in E.coli. In: FASEB Journal. 1997 ; Vol. 11, No. 9.
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