Isolation of Nine Different Biologically and Immunologically Active Molecular Variants of Bovine Follicular Inhibin1
| Autor: | James J. Ireland, Basavdutta R. Ghosh, J. Pulaski, J.L.H. Ireland, Patty S.D. Weber, Tara E.M. Good, Vasantha Padmanabhan, Alan L. Schneyer, N. Groome, G. Lambert-Messerlian, William L. Miller |
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| Rok vydání: | 1995 |
| Předmět: |
endocrine system
medicine.medical_specialty Pituitary gland biology Heterologous Biological activity Ovary Cell Biology General Medicine Molecular biology Endocrinology medicine.anatomical_structure Reproductive Medicine Affinity chromatography Immunoblot Analysis Internal medicine Electroelution medicine biology.protein Antibody hormones hormone substitutes and hormone antagonists |
| Zdroj: | Biology of Reproduction. 53:1478-1488 |
| ISSN: | 1529-7268 0006-3363 |
| Popis: | A combination of immunoaffinity chromatography, SDS-PAGE, and electroelution was used to simultaneously isolate 0.36-4.65 mg of nine different molecular forms of inhibin (proac-29 kDa; fully processed 34 kDa; and large inhibin forms 49, 53, 58, 77, 88, 110, and > 160 kDa) from 0.675 L of bovine follicularfluid (bFF). Each inhibin form, except proac, cross-reacted with inhibin ac1 -26 - and pA 82 " 4 -subunitdirected antibodies during immunoblot analysis. Proac cross-reacted only with a-subunit antibodies. The inhibin forms consisted of 22-, 29-, 49-, or 58-kDa a subunits and 17- or 58-kDa subunits. During cultures of ovine pituitary cells, a 5-ng/ml dose of each inhibin form (except proa) suppressed basal accumulation of FSH 30% to 50% but increased GnRH-induced LH release 40% to 248%. The various inhibin forms cross-reacted in a parallel fashion with standard curves generated during homologous and heterologous RIAs but with markedly different relative immunopotencies. In the RIAs, proac cross-reacted 3- to 18-fold more than the fully processed inhibin form. The fully processed and the seven different large forms of inhibin cross-reacted with different relative immunopotencies in a two-site dimer-specific ELISA. We concluded that 1) a combination of immunoaffinity extraction, SDS-PAGE, and electroelution simultaneously isolated relatively large amounts of highly enriched preparations of nine different molecular forms of immunologically and biologically active inhibin from bFF; 2) eight different dimeric forms of bovine inhibin may regulate both basal FSH and GnRH-induced LH secretion by the pituitary gland, and 3) eight or nine different molecular forms of inhibin cross-react with different relative immunopotencies in the two-site dimer-specific assay or RIAs. |
| Databáze: | OpenAIRE |
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