Macromolecular Enzymatic Product of NAD+in Liver Mitochondria

Rat liver mitochondria contain a Mg2+-requiring system that transfers the ADP-ribose moiety of NAD+to an acceptor protein. The enzyme system was extracted in a soluble form and the ADP-ribosylated protein product was isolated by hydroxyapatite and Sephadex chromatography. The ADP-ribosylated protein...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1975-04, Vol.72 (4), p.1436-1440
Main Authors: Kun, Ernest, Zimber, Peter H., Annie C. Y. Chang, Puschendorf, Bernd, Grunicke, Hans
Format: Article
Language:English
Subjects:
Adenosine Diphosphate Sugars - metabolism
Animals
Binding Sites
Chromatography
Drug Stability
Electrophoresis, Polyacrylamide Gel
Enzymes
Gels
Hydroxyapatites
Kinetics
Liver mitochondria
Mitochondria
Mitochondria, Liver - metabolism
NAD - metabolism
Nucleotides
Phosphates
Poly Adenosine Diphosphate Ribose - metabolism
Product labeling
Protein Binding
Proteins - isolation & purification
Proteins - metabolism
Radioactive decay
Radiocarbon
Rats
Receptors, Drug
Ribose - metabolism
Snake venoms
Time Factors
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Rat liver mitochondria contain a Mg2+-requiring system that transfers the ADP-ribose moiety of NAD+to an acceptor protein. The enzyme system was extracted in a soluble form and the ADP-ribosylated protein product was isolated by hydroxyapatite and Sephadex chromatography. The ADP-ribosylated protein product has a molecular weight of 100,000 and can be dissociated into subunits of 50,000 daltons by sodium dodecyl sulfate gel electrophoresis. Incubation of the isotopically labeled ADP-ribosylated protein with nicotinamide and a mitochondrial extract yields labeled NAD+, indicating apparent reversibility of the reaction. Enzymatic degradation of the ADP-ribosylated protein with snake venom phosphodiesterase liberates AMP and ADP-ribose or its isomer. Identification of these products and reversibility of the reaction show that the ADP-ribose moiety of NAD+is the molecular species that is transferred to the acceptor protein. A fraction of the protein-bound ADP-ribose appears to be present as an oligomer. The enzymatic protein-ADP-ribosylating reaction is inhibited by nicotinamide, ADP-ribose, the fluorophosphate of AMP, and picrylsulfonic acid.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.72.4.1436