PMID: 2492419Feb 1, 1989Paper

Topographic studies of microsomal and pure prostaglandin H synthase

Archives of Biochemistry and Biophysics
Richard J Kulmacz, K K Wu

Abstract

Prostaglandin H synthase catalyzes the first step in the conversion of polyunsaturated fatty acids to prostaglandins, thromboxanes, and prostacyclins. The enzyme is normally bound to the endoplasmic reticulum membrane, but can be purified to homogeneity after solubilization with detergent. The topologies of the microsomal and the pure detergent-solubilized forms of the synthase were compared by an examination of their sensitivity to degradation by proteases, of the effect of heme on this protease sensitivity, and of the sizes of proteolytic fragments produced. For the microsomal synthase, the localization of proteolytic fragments was also determined. Analysis of the microsomal proteins after proteolytic digests involved separation by polyacrylamide gel electrophoresis and selective detection of the synthase-derived polypeptides with a polyclonal antibody against the pure synthase. With both the microsomal and the pure synthase, incubation with trypsin led to a progressive loss of cyclooxygenase activity and cleavage of the synthase subunit (70K Da) into two fragments of 38K and 33K Da. Incubation of the detergent-solubilized form of the synthase with proteinase K and chymotrypsin also produced a very similar pair of fragments (...Continue Reading

References

Jan 29, 1979·Biochimica Et Biophysica Acta·F J van der OuderaaD A van Dorp
Dec 29, 1975·Biochimica Et Biophysica Acta·J W Depierre, G Dallner
May 25, 1977·Biochimica Et Biophysica Acta·F J Van der OuderaaD A Van Dorp
Mar 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·D L DeWitt, W L Smith
Jan 1, 1988·Annual Review of Biophysics and Biophysical Chemistry·G E Schulz
Mar 15, 1985·European Journal of Biochemistry·J H MutsaersJ F Vliegenthart
Jan 29, 1982·Biochemical and Biophysical Research Communications·R J Kulmacz, W E Lands
Apr 1, 1982·Archives of Biochemistry and Biophysics·B G TitusW E Lands

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Citations

Jun 20, 2003·Progress in Lipid Research·Richard J KulmaczAh-Lim Tsai
Sep 16, 1998·Current Opinion in Chemical Biology·L J Marnett, A S Kalgutkar
Mar 7, 2002·The Biochemical Journal·Hui DengKe-He Ruan
Apr 22, 2004·The Journal of Biological Chemistry·Wen LiuRichard J Kulmacz
Aug 1, 1997·Archives of Biochemistry and Biophysics·Q GuoR J Kulmacz

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