| Preliminary physico-chemical characterization of the soluble Mn2+-dependent adenylate cyclase in the rat testis. | |
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MedLine Citation:
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PMID: 7251203 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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This paper reports some physico-chemical properties of the Mn+-dependent adenylate cyclase (AC) of the rat testis. The AC activity in the crude cytosol (106 000 x g) migrates as a single peak in a linear sucrose gradient (5--20%) with a sedimentation coefficient (S20,w) of 4,1. The enzyme exhibits an Einstein-Stokes radius (rs) of 30.6 A as assessed by gel filtration (Kav = 0.50) on a calibrated Sephadex G-200 column. The molecular shape is close to spherical (f/f0 = 1.25). Isoelectric focussing of the AC peak (Kav = 0.50) from the Sephadex G-200 column revealed a pI of 5.7. Ion exchange chromatography of the crude cytosol on a Whatman DE-52 column gave a single peak eluting between 0.13--0.17 M NaCl. Electrophoresis on polyacrylamide gels resulted in a migrating AC peak with an apparent Rf of 0.40 relative to bromophenol blue. The Mn2+-dependent AC is not retained on a Con-A Sepharose 4B column indicating that the enzyme molecule does not contain sugars possessing alpha-D-manno- or alpha-D-glucopyranosyl terminal groups. (NH4)2SO4 precipitates the AC activity (4 degrees C) between 30--60% saturation. The yield approximates 85% of total AC activity with the specific activity reaching a plateau (110 pmoles cAMP/mg protein/min.) at 50% saturated (NH4)2SO4. The soluble Mn+-dependent AC of the rat testis thus appears to be relatively symmetrical with a size of approximately 52 000 D and a negative charge at physiological pH. Apparently it does not contain sugar moieties and it exhibits little molecular heterogeneity. |
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Authors:
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J O Gordeladze; D Andersen; V Hansson |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: International journal of andrology Volume: 4 ISSN: 0105-6263 ISO Abbreviation: Int. J. Androl. Publication Date: 1981 Apr |
Date Detail:
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Created Date: 1981-09-15 Completed Date: 1981-09-15 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8000141 Medline TA: Int J Androl Country: DENMARK |
Other Details:
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Languages: eng Pagination: 183-95 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adenylate Cyclase
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metabolism* Animals Centrifugation, Density Gradient Chromatography, DEAE-Cellulose Chromatography, Gel Electrophoresis, Polyacrylamide Gel Isoelectric Focusing Isoelectric Point Male Manganese / metabolism* Molecular Weight Rats Testis / enzymology* |
| Chemical | |
Reg. No./Substance:
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7439-96-5/Manganese; EC 4.6.1.1/Adenylate Cyclase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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