Document Detail


Preliminary physico-chemical characterization of the soluble Mn2+-dependent adenylate cyclase in the rat testis.
MedLine Citation:
PMID:  7251203     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
J O Gordeladze; D Andersen; V Hansson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  International journal of andrology     Volume:  4     ISSN:  0105-6263     ISO Abbreviation:  Int. J. Androl.     Publication Date:  1981 Apr 
Date Detail:
Created Date:  1981-09-15     Completed Date:  1981-09-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8000141     Medline TA:  Int J Androl     Country:  DENMARK    
Other Details:
Languages:  eng     Pagination:  183-95     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adenylate Cyclase / 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:
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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