Document Detail


Purification and properties of an (ADP-ribose)n glycohydrolase from guinea pig liver nuclei.
MedLine Citation:
PMID:  3941108     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An (ADP-ribose)n glycohydrolase has been purified more than 3,000-fold from guinea pig liver nuclei with an 18% yield. The glycohydrolase activity present in the nuclei was solubilized only by sonication at high ionic strength and purified by sequential chromatographic steps on phosphocellulose, DEAE-cellulose, Blue Sepharose, and single-stranded DNA cellulose. The purified protein exhibited one predominant protein band on sodium dodecyl sulfate-polyacrylamide gels with an estimated molecular weight of 75,500. On Sephadex G-100 gel filtration, single coincident peaks of (ADP-ribose)n glycohydrolase activity and protein with a molecular weight value of 72,000 were observed. The Km value for (ADP-ribose)n and the maximal velocity of the highly purified glycohydrolase were 2.3 microM and 36 mumol of ADP-ribose released from (ADP-ribose)n . min-1 . mg protein-1, respectively. Hydrolysis of (ADP-ribose)n by the enzyme was exoglycosidic in nature. The optimum pH for the enzyme activity was apparent at 6.8-7.0. Sulfhydryl compounds and monovalent cations were required for the maximal activity. The enzyme was sensitive to Ca2+ but not to Mg2+. The enzyme activity was inhibited by ADP-ribose, cyclic AMP (adenosine 3':5'-monophosphate) and diadenosine 5',5'''-p1,p4-tetraphosphate. Denatured DNA and histones were inhibitory, but native DNA and its histone complex were not inhibitory. Our data indicate that the glycohydrolase is present only as a minor protein in nuclei, being present in perhaps about 50,000 molecules/nucleus.
Authors:
S Tanuma; K Kawashima; H Endo
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  261     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1986 Jan 
Date Detail:
Created Date:  1986-02-14     Completed Date:  1986-02-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  965-9     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cations / pharmacology
Cell Nucleus / enzymology*
Chromatography, DEAE-Cellulose
Chromatography, Gel
DNA / pharmacology
Electrophoresis, Polyacrylamide Gel
Glycoside Hydrolases / isolation & purification*
Guinea Pigs
Histones / pharmacology
Hydrogen-Ion Concentration
Kinetics
Liver / cytology*
Sonication
Sulfhydryl Compounds / metabolism
Chemical
Reg. No./Substance:
0/Cations; 0/Histones; 0/Sulfhydryl Compounds; 9007-49-2/DNA; EC 3.2.1.-/Glycoside Hydrolases; EC 3.2.1.143/poly ADP-ribose glycohydrolase

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