[Enzymatic synthesis of beta-NAD+ selectively marked with tritium at adenine and its use for determining the activity of poly(ADP-ribose) polymerase].
A method of preparation of tritiated beta-HA [symbol: see text]+ from (adenine-2,8-(3)H)ATP and nicotinamide mononucleotide is suggested. This method is based on the baker's yeast (Saccharomyces cerevisiae) enzyme nicotinamide nucleotide adenylyltransferase. Experimental conditions for a nearly complete conversion of labeled ATP to NAD were determined. The tritiated NAD prepared by this method can be used in the quantitative assay of the chromatin enzyme poly(ADP-ribose) polymerase.
Shram
S I
SI
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia. img@glas.apc.org
Rybakova
I G
IG
Lazurkina
T Iu
TIu
Sidorov
G V
GV
Shilovskiĭ
G A
GA
Khokhlov
A N
AN
Miasoedov
N F
NF
D000225:Adenine / Q000737:chemistry*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D015335:Molecular Probes; D009243:NAD / Q000138:chemical synthesis / Q000737:chemistry*; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D014316:Tritium
D015335:Molecular Probes; D009243:NAD; D014316:Tritium; D011065:Poly(ADP-ribose) Polymerases; D009612:Nicotinamide-Nucleotide Adenylyltransferase; D000225:Adenine