ABSTRACT

The key metabolite of vitamin D, 1,25-dihydroxyvitamin D (1,25-D), induces rapid cellular responses that constitute a so-called ``non-genomic'' response. This effect is distinguished from its ``classic'' genomic role in calcium homeostasis involving the nuclear 1,25-D receptor. Evidence is presented that protein kinase C (PKC) is directly activated by 1,25-D at physiological concentrations (EC = 16 ± 1 nM). The effect was demonstrable with single PKC-, -, and - isoform preparations, assayed in a system containing only purified enzyme, substrate, co-factors, and lipid vesicles, from which it is inferred that a direct interaction with the enzyme is involved. The finding that calcium-independent isoform PKC- was also activated by 1,25-D shows that the calcium binding C2 domain is not required. The level of 1,25-D-induced activation, paired with either diacylglycerol or 4-12-O-tetradecanoylphorbol-13-acetate, was greater than that achievable by any individual activator alone, each at a saturating concentration, a result that implies two distinct activator sites on the PKC molecule. Phosphatidylethanolamine present in the lipid vesicles potentiated 4-12-O-tetradecanoylphorbol-13-acetate- and diacylglycerol-induced PKC activities, whereas 1,25-D-induced activity decreased, consistent with 1,25-D-activated PKC possessing a distinct conformation. The results suggest that PKC is a ``membrane-bound receptor'' for 1,25-D and that it could be important in the control of non-genomic cellular responses to the hormone.

FOOTNOTES

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This work was supported by U. S. Public Health Service Grants AA08022, AA07215, AA07186, and AA07465. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked ``advertisement'' in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§

To whom correspondence should be addressed: Dept. of Pathology and Cell Biology, Rm. 271 JAH, Thomas Jefferson University, Philadelphia, PA 19107. Tel.: 215-955-5019; stubbsc{at}jeflin.tju.edu.

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The abbreviations used are: 1,25-D, 1,25-dihydroxyvitamin D; PKC, protein kinase C; LUV, large unilamellar vesicles; TPA, 4-12-O-tetradecanoylphorbol-13-acetate; DAG, diacylglycerol; PE, phosphatidylethanolamine; BPS, brain phosphatidylserine; POPC, palmitoyl-oleoylphosphatidylcholine.

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M. D. Yeager, F. J. Taddeo, S. J. Slater, and C. D. Stubbs, manuscript in preparation.

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S. J. Slater, M. B. Kelly, and C. D. Stubbs, unpublished observations.

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