Alternative Splicing Determines the Function of CYP4F3 by Switching Substrate Specificity

Peter Christmas, Jeffrey P. Jones, Christopher J. Patten, Dan A. Rock, Yimin Zheng, Shing Ming Cheng, Brittany M. Weber, Nadia Carlesso, David T. Scadden, Allan E. Rettie, Roy J. Soberman

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Abstract

Diversity of cytochrome P450 function is determined by the expression of multiple genes, many of which have a high degree of identity. We report that the use of alternate exons, each coding for 48 amino acids, generates isoforms of human CYP4F3 that differ in substrate specificity, tissue distribution, and biological function. Both isoforms contain a total of 520 amino acids. CYP4F3A, which incorporates exon 4, inactivates LTB4 by ω-hydroxylation (Km = 0.68 μM) but has low activity for arachidonic acid (K m = 185 μM); it is the only CYP4F isoform expressed in myeloid cells in peripheral blood and bone marrow. CYP4F3B incorporates exon 3 and is selectively expressed in liver and kidney; it is also the predominant CYP4F isoform in trachea and tissues of the gastrointestinal tract. CYP4F3B has a 30-fold higher Km for LTB4 compared with CYP4F3A, but it utilizes arachidonic acid as a substrate for ω-hydroxylation (K m = 22 μM) and generates 20-HETE, an activator of protein kinase C and Ca2+/calmodulin-dependent kinase II. Homology modeling demonstrates that the alternative exon has a position in the molecule which could enable it to contribute to substrate interactions. The results establish that tissue-specific alternative splicing of pre-mRNA can be used as a mechanism for changing substrate specificity and increasing the functional diversity of cytochrome P450 genes.

Original languageEnglish (US)
Pages (from-to)38166-38172
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number41
StatePublished - Oct 12 2001

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Christmas, P., Jones, J. P., Patten, C. J., Rock, D. A., Zheng, Y., Cheng, S. M., Weber, B. M., Carlesso, N., Scadden, D. T., Rettie, A. E., & Soberman, R. J. (2001). Alternative Splicing Determines the Function of CYP4F3 by Switching Substrate Specificity. Journal of Biological Chemistry, 276(41), 38166-38172.