Expression and Functional Characterization of Breast Cancer-Associated Cytochrome P450 4Z1 in Saccharomyces cerevisiae .

Autor: McDonald MG; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Ray S; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Amorosi CJ; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Sitko KA; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Kowalski JP; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Paco L; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Nath A; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Gallis B; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Totah RA; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Dunham MJ; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Fowler DM; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington., Rettie AE; Departments of Medicinal Chemistry (M.G.M., S.R., J.P.K., L.P., A.N., B.G., R.A.T., A.E.R.), Genome Sciences (K.A.S., C.J.A., M.J.D., D.M.F.), and Bioengineering (D.M.F.), University of Washington, Seattle, Washington rettie@uw.edu.
Jazyk: angličtina
Zdroj: Drug metabolism and disposition: the biological fate of chemicals [Drug Metab Dispos] 2017 Dec; Vol. 45 (12), pp. 1364-1371. Date of Electronic Publication: 2017 Oct 10.
DOI: 10.1124/dmd.117.078188
Abstrakt: CYP4Z1 is an "orphan" cytochrome P450 (P450) enzyme that has provoked interest because of its hypothesized role in breast cancer through formation of the signaling molecule 20-hydroxyeicosatetraenoic acid (20-HETE). We expressed human CYP4Z1 in Saccharomyces cerevisiae and evaluated its catalytic capabilities toward arachidonic and lauric acids (AA and LA). Specific and sensitive mass spectrometry assays enabled discrimination of the regioselectivity of hydroxylation of these two fatty acids. CYP4Z1 generated 7-, 8-, 9-, 10-, and 11-hydroxy LA, whereas the 12-hydroxy metabolite was not detected. HET0016, the prototypic CYP4 inhibitor, only weakly inhibited laurate metabolite formation (IC 50 ∼15 μ M). CYP4Z1 preferentially oxidized AA to the 14(S),15(R)-epoxide with high regioselectivity and stereoselectivity, a reaction that was also insensitive to HET0016, but neither 20-HETE nor 20-carboxy-AA were detectable metabolites. Docking of LA and AA into a CYP4Z1 homology model was consistent with this preference for internal fatty acid oxidation. Thus, human CYP4Z1 has an inhibitor profile and product regioselectivity distinct from most other CYP4 enzymes, consistent with CYP4Z1's lack of a covalently linked heme. These data suggest that, if CYP4Z1 modulates breast cancer progression, it does so by a mechanism other than direct production of 20-HETE.
(Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.)
Databáze: MEDLINE
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