Identification and purification of a bovine liver mitochondrial NAD(+)-glycohydrolase

FEBS Lett. 1995 Dec 27;377(3):530-4. doi: 10.1016/0014-5793(95)01366-0.


Nonenzymatic ADP-ribosylation of mitochondrial proteins is thought to play a role in the regulation of Ca2+ efflux from mitochondria. It has been shown that intramitochondrial ADP-ribose is generated by a specific NAD(+)glycohydrolase, which catalizes hydrolysis of NAD+ to ADP-ribose and nicotinamide. We purified this enzyme from bovine liver mitochondrial membranes. The final preparation had a 1660-fold purified enzyme activity and contained a main protein band with an apparent molar mass of 32,000 in a SDS-polyacrylamide gel. The identity of this protein band with NAD(+)-glycohydrolase was verified by renaturation of its enzymatic activity. Partial amino acid sequence information was obtained from two enzyme fragments after proteolytic cleavage of the protein band in the SDS-polyacrylamide gel. Searches in protein databases revealed that an arginine ADP-ribosyl hydrolase harbours two stretches of amino acids that are highly similar to the partial NAD(+)-glycohydrolase sequences.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cattle
  • Chromatography
  • Electrophoresis, Polyacrylamide Gel
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism
  • Mitochondria, Liver / enzymology*
  • Molecular Sequence Data
  • NAD+ Nucleosidase / chemistry
  • NAD+ Nucleosidase / isolation & purification*
  • NAD+ Nucleosidase / metabolism
  • Protein Folding
  • Sequence Analysis
  • Submitochondrial Particles / enzymology


  • Membrane Proteins
  • NAD+ Nucleosidase