HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Murphy, R. C. Articles by Schwartzman, M. L. Search for Related Content PubMed PubMed Citation Articles by Murphy, R. C. Articles by Schwartzman, M. L. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 263, Issue 32, 17197-17202, 11, 1988

12(R)-hydroxyeicosatrienoic acid: a vasodilator cytochrome P-450- dependent arachidonate metabolite from the bovine corneal epithelium

RC Murphy, JR Falck, S Lumin, P Yadagiri, JA Zirrolli, M Balazy, JL Masferrer, NG Abraham and ML Schwartzman
Department of Pharmacology, University of Colorado Health Science Center, Denver 80262.

When corneal microsomes were incubated with arachidonic acid in the presence of an NADPH-generating system, two biologically active metabolites of arachidonic acid were formed. The structure of one of the metabolites, compound C, was previously reported to be 12(R)- hydroxy-5,8,10,14-eicosatetraenoic acid and was found to be a potent inhibitor of the Na+/K+-ATPase in the cornea. The second metabolite, compound D, was found to be a potent vasodilator as well as having the property of stimulating protein influx into the aqueous humor of the eye. Following purification of compound D by thin layer chromatography and high pressure liquid chromatography, it was found to lack a UV chromophore in contrast to the previously reported cytochrome P-450- dependent metabolite. Mass spectrometric analysis using positive and negative ionization modes was carried out on derivatized compound D that had been synthesized from a mixture of labeled [( 5,6,8,9,11,12,14,15-2H8]) and unlabeled arachidonic acid incubated with corneal microsomes. The novel arachidonate metabolite had abundant fragment ions consistent with compound D being a monooxygenated derivative of arachidonic acid with a hydroxyl substituent at carbon 12 of the eicosanoid backbone; only seven deuterium atoms from [2H8]arachidonate were retained in the structure. Oxidative ozonolysis yielded a product indicating that the double bonds in metabolite D resided between carbons at positions 8 and 9 and positions 14 and 15 of the 20-carbon chain. Compound D was therefore characterized as 12- hydroxy-5,8,14-eicosatrienoic acid. Model compounds were synthesized from dimethyl malate with the hydroxy at the 12 position with both the R and S absolute configuration and with all double bonds of the cis configuration. Only the 12(R) isomer was found to be a potent vasodilator and to increase aqueous humor protein concentration, suggesting that the biologically active compound D was 12(R)-hydroxy- 5,8,14-(Z,Z,Z)-eicosatrienoic acid. As this compound possesses proinflammatory properties, it may play a role in the wound-healing processes of corneal injury.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L. Du, V. Yermalitsky, D. L. Hachey, S. G. Jagadeesh, J. R. Falck, and D. S. Keeney A Biosynthetic Pathway Generating 12-Hydroxy-5,8,14-eicosatrienoic Acid from Arachidonic Acid Is Active in Mouse Skin Microsomes J. Pharmacol. Exp. Ther., January 1, 2006; 316(1): 371 - 379. [Abstract] [Full Text] [PDF]

				P. A. Mieyal, M. W. Dunn, and M. L. Schwartzman Detection of Endogenous 12-Hydroxyeicosatrienoic Acid in Human Tear Film Invest. Ophthalmol. Vis. Sci., February 1, 2001; 42(2): 328 - 332. [Abstract] [Full Text]

				J. H. Capdevila, J. R. Falck, and R. C. Harris Cytochrome P450 and arachidonic acid bioactivation: molecular and functional properties of the arachidonate monooxygenase J. Lipid Res., February 1, 2000; 41(2): 163 - 181. [Abstract] [Full Text]

				M. K. Patricia, J. A. Kim, C. M. Harper, P. T. Shih, J. A. Berliner, R. Natarajan, J. L. Nadler, and C. C. Hedrick Lipoxygenase Products Increase Monocyte Adhesion to Human Aortic Endothelial Cells Arterioscler. Thromb. Vasc. Biol., November 1, 1999; 19(11): 2615 - 2622. [Abstract] [Full Text] [PDF]

				V. Mastyugin, E. Aversa, A. Bonazzi, C. Vafaes, P. Mieyal, and M. L. Schwartzman Hypoxia-Induced Production of 12-Hydroxyeicosanoids in the Corneal Epithelium: Involvement of a Cytochrome P-4504B1 Isoform J. Pharmacol. Exp. Ther., June 1, 1999; 289(3): 1611 - 1619. [Abstract] [Full Text]