PMID: 3766957Aug 15, 1986Paper

Effect of photoaffinity labeling on rabbit uterine progesterone receptor

Analytical Biochemistry
C L ClarkeP G Satyaswaroop

Abstract

Photoaffinity labeling with [17 alpha-methyl-3H]promegestone ([ 3H]R5020) is an effective technique for the covalent labeling of the progesterone receptor (PR), which allows monitoring of the steroid receptor complex under denaturing conditions. The present study was initiated to evaluate whether photolabeled PR could be used also as a marker for PR under nondenaturing conditions. Accordingly, the effect of irradiation on each component of the reaction was evaluated separately. When [3H]R5020 alone was irradiated, there was a rapid (less than 5 min), light dependent destruction of [3H]R5020, as evident from increased formation of a more polar tritiated product on TLC and a concomitant decrease in the ability of the irradiated preparation to bind to PR. When rabbit uterine PR was irradiated in the absence of steroid, a gradual decrease in the binding capacity was observed, reaching 70% of the nonirradiated control in 10 min. The optimal irradiation time for covalent [3H]R5020-PR complex formation was determined by irradiation for up to 5 min, and separation of the products by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis. Specific labeling of proteins of Mr 116,000 and 85,000 was observed, with the rate of label...Continue Reading

References

Mar 28, 1977·Biochimica Et Biophysica Acta·O VesterbergA Sjösten
Jan 1, 1985·Canadian Journal of Biochemistry and Cell Biology = Revue Canadienne De Biochimie Et Biologie Cellulaire·P K Tai, L E Faber
Aug 1, 1984·Molecular and Cellular Endocrinology·J M Renoir, J Mester
Sep 1, 1981·European Journal of Biochemistry·H M WestphalM Beato

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Citations

Feb 1, 1990·Journal of Steroid Biochemistry·K G PinneyJ A Katzenellenbogen
Mar 1, 1991·The Journal of Steroid Biochemistry and Molecular Biology·W SchneiderG Shyamala

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