Author:
Khan F S,Rathnam P,Saxena B B
Abstract
Plasma-membrane and soluble fractions containing human chorionic gonadotropin/lutropin receptor were prepared from bovine corpora lutea by ultracentrifugation. The plasma-membrane and soluble fractions were studied for physicochemical properties, salts and gangliosides. The receptor preparations obtained from the plasma-membrane purified individually by sucrose-density-gradient centrifugation, which resulted in a partial dissociation of the hormone-binding subunit from the intact functional receptor unit, which consists of both hormone-binding (regulatory) and adenylate cyclase-associated (catalytic) subunits. The fractions containing the functional receptor unit were further purified by gel filtration on Sepharose-6B and chromatography on concanavalin A-Sepharose. The ‘receptor’ was finally purified by affinity chromatography on a column of controlled-pore glass covalently coupled to hu man chorionic gonadotropin. The purified receptor from the plasma-membrane and the soluble fractions contained binding capacities of 901000 and 87000 fmol of human chorionic gonadotropin/mg of protein. Yields of 0.02 and 0.22mg of protein were obtained from 250 g of bovine corpora lutea, which represents a 10000- and 1000-fold increase respectively in the specific binding with 125I-labelled human chorionic gonadotropin. Immunization of rabbits with a partially purified receptor fraction generated antibodies that specifically inhibited the binding of the 125I-labelled human chorionic gonadotropin to the receptor.
Subject
Cell Biology,Molecular Biology,Biochemistry
Cited by
11 articles.
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1. Expression of a recombinant bifunctional protein from a chimera of human lutropin receptor and human chorionic gonadotropin β-subunit;Journal of Reproductive Immunology;2004-10
2. Human corpus luteum: human chorionic gonadotropin receptors during ectopic pregnancy**Presented in part at the 47th Annual Meeting of The American Fertility Society, Orlando, Florida, October 21 to 24, 1991.;Fertility and Sterility;1994-10
3. Luteinizing hormone and human chorionic gonadotropin receptors in human corpora lutea from clomiphene citrate-induced cycles**Presented in part at the 36th Annual Meeting of the Society for Gynecologic Investigation, San Diego, California, March 15 to 18, 1989. This work was supported by an American College of Obstetricians and Gynecologists-Ortho Fellowship Grant and ACOG District VI, Illinois Section, Research Grant.;Fertility and Sterility;1990-10
4. The Follicle-Stimulating Hormone (FSH) Receptor in Testis: Interaction with FSH, Mechanism of Signal Transduction, and Properties of the Purified Receptor1;Biology of Reproduction;1989-01-01
5. Molecular Structure of the Luteinizing Hormone Receptor1;Biology of Reproduction;1989-01-01