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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
The Effect of Leupeptin on Progestin Receptors of Human Uterine Tissues
Journal of Clinical Endocrinology and Metabolism, Volume 55, No. 5, Year 1982
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Description
In preparation for studies of progestin receptor dynamics in human uterine tissues, we have investigated the effect of tissue handling and the protease inhibitor leupeptin on the molecular forms of progestin receptors in human endometrium, myometrium, and leiomyoma. Tritiated R5020 [17,21–dimethyl–19–nor–4,9–pregnadiene–3,20–dione) (promegestone)] was the labeled ligand, and sucrose density gradient ultracentrifugation was used to determine the sedimentation coefficients of the receptors. Scatchard plots of saturation analyses were used to determine the number of binding sites and dissociation constants. We found that rapid chilling of tissue at surgery was necessary to maintain specific progestin binding in myometrium, that leupeptin permitted demonstration of 8S receptors in my-ometrium and leiomyoma, and that 8S receptors were present in endometrium, with or without leupeptin. With careful handling of tissue and minimal homogenization, leupeptin may not be necessary to preserve 8S receptors in myometrium and leiomy-oma cytosols; however, the use of leupeptin gives greater assurance that 8S receptors will be preserved. The number of binding sites varied from 0.6–2.0 pmol⁄mg cytosol protein, and the Kd values observed were between 1.7–6.9 × 10−10 M. © 1982 by The Endocrine Society.
Authors & Co-Authors
Eldieb, Mohammad R.
United States, Memphis
University of Tennessee College of Medicine Memphis
Egypt, Cairo
Faculty of Medicine
Andersen, Richard N.
United States, Memphis
University of Tennessee College of Medicine Memphis
Statistics
Citations: 8
Authors: 2
Affiliations: 2
Identifiers
Doi:
10.1210/jcem-55-5-1007
ISSN:
0021972X
e-ISSN:
19457197
Research Areas
Health System And Policy
Study Approach
Qualitative