A MODIFICATION TO AN ENZYMATIC METHOD FOR THE PREPARATION OF (ALPHA-P-32) ATP WITH A HIGH SPECIFIC ACTIVITY

Authors
Citation
F. Wu et al., A MODIFICATION TO AN ENZYMATIC METHOD FOR THE PREPARATION OF (ALPHA-P-32) ATP WITH A HIGH SPECIFIC ACTIVITY, Journal of radioanalytical and nuclear chemistry, 218(1), 1997, pp. 41-43
Citations number
7
Categorie Soggetti
Chemistry Inorganic & Nuclear","Chemistry Analytical","Nuclear Sciences & Tecnology
ISSN journal
02365731
Volume
218
Issue
1
Year of publication
1997
Pages
41 - 43
Database
ISI
SICI code
0236-5731(1997)218:1<41:AMTAEM>2.0.ZU;2-F
Abstract
A modification of the procedure reported by JOHNSON and WALSETH(1) for the preparation of (alpha-P-32)ATP : (gamma-P-32)GTP with a specific activity of more than 5000 Ci/mmol was made firstly, then it was used as the phosphate donor in the reaction catalyzed by the polynucleotide kinase, to transfer its radioactive phosphate to the 5'-position of t he 3'-AMP, resulting (5'-P-32)3'-ADP. After that, (5'-P-32)3'-ADP was hydrolysed to (5'-P-32)AMP under the catalysis of Nuclease P1, the lat e was then phosphorylated to produce (alpha-P-32)ATP by the action of myokinase and pyruvate kinase. By this method, we firstly prepared (al pha-P-32)ATP with a specific activity of more than 4000 Ci/mmol which was measured by HPLC analysis.