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Identification and purification of a bovine liver mitochondrial NAD + ‐glycohydrolase
Author(s) -
Ju Zhang,
Mathias Ziegler,
Rainer Schneider,
Helmut Klocker,
Bernhard Auer,
Manfred Schweiger
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01366-0
Subject(s) - nad+ kinase , chemistry , identification (biology) , biochemistry , microbiology and biotechnology , biology , enzyme , botany
Nonenzymatic ADP‐ribosylation of mitochondrial proteins is thought to play a role in the regulation of Ca 2+ 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.