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.
Subject(s)
Adenine/chemistry , NAD/chemistry , Poly(ADP-ribose) Polymerases/metabolism , Animals , Base Sequence , Cell Line , Cricetinae , Cricetulus , Molecular Probes , NAD/chemical synthesis , Nicotinamide-Nucleotide Adenylyltransferase/metabolism , Saccharomyces cerevisiae/enzymology , TritiumABSTRACT
Formation of prostaglandins A, B and F from prostaglandin E has been studied. Synthesis of tritium-labelled prostaglandins A1, A2, B1, B2, F1 alpha, F2 alpha, F1 beta, F2 beta of high molar radioactivity from highly labelled PGE1 and PGE2 is described.
Subject(s)
Prostaglandins A, Synthetic , Prostaglandins B , Prostaglandins F, Synthetic , Prostaglandins, Synthetic , Prostaglandins , Chemical Phenomena , Chemistry , TritiumABSTRACT
By selecting specific activators of PGF reductase and determining their optimal ratio, we have performed the directed biosynthesis of PGF2 alpha with a 45-50% yield. With the use of [5,6,8,9,11,12,14,15-3H8]arachidonic acid, multiple-labelled PGF2 alpha with molar radioactivity 6.1 TBe/mmol has been synthesised.
