PMID: 6304691May 1, 1983Paper

Specific labeling of mouse brain membrane phospholipids with [20-3H]phorbol 12-p-azidobenzoate 13-benzoate, a photolabile phorbol ester

Proceedings of the National Academy of Sciences of the United States of America
K B DelclosP M Blumberg

Abstract

As part of our effort to characterize receptors for the phorbol ester tumor promoters, a phorbol ester photoaffinity probe, [20-3H]phorbol 12-p-azidobenzoate 13-benzoate (PaBzBz), was synthesized. In the dark, PaBzBz bound reversibly to brain particulate fractions with a dissociation constant (Kd) of 0.81 +/- 0.09 x 10(-9) M. Specific binding of PaBzBz, at a concentration equal to its Kd, represented 85% of the total bound. At saturation, 24 +/- 5 pmol of PaBzBz were bound per mg of brain protein, a level similar to that observed with [20-3H]phorbol 12,13-dibutyrate. Under the conditions used (concentrations greater than the Kd for PaBzBz), irradiation caused 45% of the PaBzBz binding to become irreversible. Most of the binding (approximately equal to 60%), including most of the specific irreversible binding, was to phospholipid rather than to protein. Based on susceptibility to enzymatic digestion and on chromatographic mobility, the specifically labeled phospholipids were identified as phosphatidylserine, phosphatidylethanolamine, and phosphatidylethanolamine plasmalogen. Although the PaBzBz specifically labeled lipid, labeling was blocked by pretreatment of membranes at 100 degrees C for 5 min or by papain digestion. Therefo...Continue Reading

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Citations

Sep 8, 1998·Bioorganic & Medicinal Chemistry·K UotsuM Shibasaki
Jul 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·K L LeachP M Blumberg
Jan 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·N A SharkeyP M Blumberg
Jan 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·A M Jeffrey, R M Liskamp
Mar 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·B R GanongR M Bell
Oct 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·P A WenderP M Blumberg
Jun 13, 2013·Journal of Cancer Research and Clinical Oncology·Hirota FujikiMasami Suganuma
Dec 5, 1984·Biochimica Et Biophysica Acta·T A BerkhoutJ J Sixma
Jul 10, 1989·Biochimica Et Biophysica Acta·K A Fisher, K C Yanagimoto
Jan 1, 1985·Journal of Cellular Biochemistry·B KönigP M Blumberg
Jul 1, 1991·Japanese Journal of Cancer Research : Gann·S NishiwakiM Natsume

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