Characterization of an additional splice acceptor site introduced into CYP4B1 in hominoidae during evolution

Eva M. Schmidt, Constanze Wiek, Oliver T. Parkinson, Katharina Roellecke, Marcel Freund, Michael Gombert, Nadine Lottmann, Charles A. Steward, Christof M. Kramm, Vladimir Yarov-Yarovoy, Allan E. Rettie, Helmut Hanenberg

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

CYP4B1 belongs to the cytochrome P450 family 4, one of the oldest P450 families whose members have been highly conserved throughout evolution. The CYP4 monooxygenases typically oxidize fatty acids to both inactive and active lipid mediators, although the endogenous ligand(s) is largely unknown. During evolution, at the transition of great apes to humanoids, the CYP4B1 protein acquired a serine instead of a proline at the canonical position 427 in the meander region. Although this alteration impairs P450 function related to the processing of naturally occurring lung toxins, a study in transgenic mice suggested that an additional serine insertion at position 207 in human CYP4B1 can rescue the enzyme stability and activity. Here, we report that the genomic insertion of a CAG triplet at the intron 5-exon 6 boundary in human CYP4B1 introduced an additional splice acceptor site in frame. During evolution, this change occurred presumably at the stage of Hominoidae and leads to two major isoforms of the CYP4B1 enzymes of humans and great apes, either with or without a serine 207 insertion (insSer207). We further demonstrated that the CYP4B1 enzyme with insSer207 is the dominant isoform (76%) in humans. Importantly, this amino acid insertion did not affect the 4-ipomeanol metabolizing activities or stabilities of the native rabbit or human CYP4B1 enzymes, when introduced as transgenes in human primary cells and cell lines. In our 3D modeling, this functional neutrality of insSer207 is compatible with its predicted location on the exterior surface of CYP4B1 in a flexible side chain. Therefore, the Ser207 insertion does not rescue the P450 functional activity of human CYP4B1 that has been lost during evolution.

Original languageEnglish (US)
Article numbere0137110
JournalPLoS One
Volume10
Issue number9
DOIs
StatePublished - Sep 10 2015

ASJC Scopus subject areas

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

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    Schmidt, E. M., Wiek, C., Parkinson, O. T., Roellecke, K., Freund, M., Gombert, M., Lottmann, N., Steward, C. A., Kramm, C. M., Yarov-Yarovoy, V., Rettie, A. E., & Hanenberg, H. (2015). Characterization of an additional splice acceptor site introduced into CYP4B1 in hominoidae during evolution. PLoS One, 10(9), [e0137110]. https://doi.org/10.1371/journal.pone.0137110