[Enzymatic synthesis of beta-NAD+ selectively marked with tritium at adenine and its use for determining the activity of poly(ADP-ribose) polymerase]

S I Shram; I G Rybakova; T Iu Lazurkina; G V Sidorov; G A Shilovskiĭ; A N Khokhlov; N F Miasoedov

[Enzymatic synthesis of beta-NAD+ selectively marked with tritium at adenine and its use for determining the activity of poly(ADP-ribose) polymerase]

Prikl Biokhim Mikrobiol. 1999 Nov-Dec;35(6):638-46.

[Article in Russian]

Authors

S I Shram¹, I G Rybakova, T Iu Lazurkina, G V Sidorov, G A Shilovskiĭ, A N Khokhlov, N F Miasoedov

Affiliation

¹ Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia. img@glas.apc.org

PMID: 10623965

Abstract

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.

MeSH terms

Adenine / chemistry*
Animals
Base Sequence
Cell Line
Cricetinae
Cricetulus
Molecular Probes
NAD / chemical synthesis
NAD / chemistry*
Nicotinamide-Nucleotide Adenylyltransferase / metabolism
Poly(ADP-ribose) Polymerases / metabolism*
Saccharomyces cerevisiae / enzymology
Tritium

Substances

Molecular Probes
NAD
Tritium
Poly(ADP-ribose) Polymerases
Nicotinamide-Nucleotide Adenylyltransferase
Adenine