Photolabeling of a O2-. generating protein in bovine polymorphonuclear neutrophils by an arylazido NADP+ analog

Biochemical and Biophysical Research Communications
J DoussiereP V Vignais

Abstract

A plasma membrane fraction of bovine polymorphonuclear neutrophils enriched in NADPH-dependent, O2-. generating oxidase activity, and a number of fractions solubilized in detergent, recovered during the course of the purification of this oxidase have been tested for their ability to react with radiolabeled N-4-azido-2-nitrophenyl aminobutyryl NADP+ (arylazido NADP+ or NAP4-NADP+). In the dark, NAP4-NADP+ and its reduced form NAP4-NADPH, were found to inhibit competitively the NADPH-dependent O2-. generating oxidase activity of the plasma membrane fraction of bovine neutrophils activated by phorbol myristate acetate. The nitrene derivative formed upon photoirradiation of NAP4-NADP(H) bound covalently to different proteins of the plasma membrane. Photolabeling of these proteins was prevented by preincubation with an excess of NADPH. Photolabeling of a protein of 65,000 Mr was decreased by omission of phorbol myristate acetate as activating agent of the respiratory burst in neutrophils or by addition of micromolar amounts of Cibacron Blue and mersalyl which are known to inhibit the production of O2-. by the plasma membrane fraction. During the course of the purification procedure, the 65000 Mr protein emerged as the preferentially...Continue Reading

References

Nov 15, 1973·Journal of Molecular Biology·U K Laemmli, M Favre

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Citations

Dec 1, 1988·Journal of Bioenergetics and Biomembranes·J F Parkinson, T G Gabig
Sep 3, 1990·Biochimica Et Biophysica Acta·Y Nisimoto, H Otsuka-Murakami
Jan 1, 1988·Free Radical Biology & Medicine·P Bellavite
Jan 1, 1989·Critical Reviews in Biochemistry and Molecular Biology·J K Hurst, W C Barrette
Jan 1, 1993·Critical Reviews in Clinical Laboratory Sciences·M C Dinauer
Jul 18, 1988·FEBS Letters·J Doussière, P V Vignais
Dec 31, 1991·Biochemical and Biophysical Research Communications·P Ravel, F Lederer
May 6, 1991·Biochimica Et Biophysica Acta·A R Cross, O T Jones

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