Document Detail


Purification, characterization and cellular localization of 5'-nucleotidase from Torpedo electric organ.
MedLine Citation:
PMID:  2822031     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
5'-Nucleotidase was isolated from the electric organ of the electric ray Torpedo marmorata after solubilization in Triton X-100 and deoxycholate by affinity chromatography on concanavalin A-Sepharose and AMP-Sepharose. The purified enzyme has a Km for AMP of 38 microM, with a maximal velocity of 31 units/mg of protein. Of the purine and pyrimidine mononucleotides, AMP is hydrolysed most effectively. beta-Glycerophosphate, phosphoenolpyruvate and p-nitrophenyl phosphate are not substrates for the enzyme. Adenosine 5'-[alpha, beta-methylene]diphosphate, ADP and ATP are competitive inhibitors in this order of potency. Concanavalin A inhibits enzyme activity in a non-competitive manner. Whereas Mg2+, Ca2+ and Sr2+ activate enzyme activity in the millimolar range, Hg2+, and in particular Pb2+ and Zn2+, inhibit enzyme activity. On SDS/polyacrylamide-gel electrophoresis the enzyme has an apparent Mr of 62000, whereas that of the native deoxycholate-enzyme complex is 131000. An antiserum raised against the native enzyme inhibits enzyme activity. Inhibition studies suggest the presence of tissue-specific variants of the enzyme. By immunohistochemical analysis the enzyme can be localized to the ramifications of nerve terminals in the electric organ.
Authors:
E J Grondal; H Zimmermann
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Biochemical journal     Volume:  245     ISSN:  0264-6021     ISO Abbreviation:  Biochem. J.     Publication Date:  1987 Aug 
Date Detail:
Created Date:  1987-11-18     Completed Date:  1987-11-18     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  2984726R     Medline TA:  Biochem J     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  805-10     Citation Subset:  IM    
Affiliation:
AK Neurochemie, Zoologisches Institut der J.W. Goethe-Universitt, Frankfurt am Main, Federal Republic of Germany.
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MeSH Terms
Descriptor/Qualifier:
5'-Nucleotidase
Animals
Cations, Divalent / pharmacology
Electric Organ / enzymology*
Electrophoresis, Polyacrylamide Gel
Fluorescent Antibody Technique
Histocytochemistry
Immune Sera / pharmacology
Nucleotidases / antagonists & inhibitors,  isolation & purification,  metabolism*
Substrate Specificity
Tissue Distribution
Torpedo / metabolism*
Chemical
Reg. No./Substance:
0/Cations, Divalent; 0/Immune Sera; EC 3.1.3.-/Nucleotidases; EC 3.1.3.5/5'-Nucleotidase
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