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PMID: 6307269 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A simple assay method for determination of the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP.

The Biochemical journal ·Vol. 210 ·No. 3 ·1983-03-15 ·Pages 717-20

Hawkins PT, Michell RH, Kirk CJ

Abstract

A simple and rapid assay method is described for determining the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP. Labelled ATP is incubated with cyclic AMP, cyclic AMP-dependent protein kinase and histone H2A under conditions leading to maximum phosphorylation of the histone. The specific radioactivity of the gamma-phosphate group of the 32P-labelled ATP is calculated from the total amount of [32P]Pi incorporated into a standard amount of histone. This assay method uses inexpensive commercially available materials, and it yields an accurate specific radioactivity with as little as 0.25 nmol of 32P-labelled ATP.

MeSH Terms

Adenosine Triphosphate/metabolism Cyclic AMP/metabolism Histones/metabolism Liver/cytology,metabolism Methods Phosphorus Radioisotopes Protein Kinases/metabolism

Chemicals

Histones Phosphorus Radioisotopes Adenosine Triphosphate Cyclic AMP Protein Kinases

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Hawkins P T

Michell R H

Kirk C J

References (6)

6 references, click to expand

Assay of cyclic AMP-dependent protein kinases.
Methods Enzymol. 1974;38:287-90 PMID: 4375761
A rapid method for the measurement of [gamma-32P]ATP specific radioactivity in tissue extracts and its application to the study of 32Pi uptake in perfused rat heart.
Anal Biochem. 1976 Oct;75(2):429-35 PMID: 984403
The role of calcium ion as a mediator of the effects of angiotensin II, catecholamines, and vasopressin on the phosphorylation and activity of enzymes in isolated hepatocytes.
J Biol Chem. 1979 Aug 10;254(15):7147-56 PMID: 222757
An easy and rapid method for the measurement of [gamma-32P]ATP specific radioactivity in tissue extracts obtained from in vitro rat heart preparations labeled with 32Pi.
Anal Biochem. 1981 Jan 15;110(2):331-7 PMID: 7235220
Phosphatidylinositol metabolism in rat hepatocytes stimulated by vasopressin.
Biochem J. 1981 Jan 15;194(1):155-65 PMID: 7030316
PURIFICATION AND PROPERTIES OF RABBIT SKELETAL MUSCLE PHOSPHORYLASE B KINASE.
Biochemistry. 1964 Aug;3:1022-33 PMID: 14220660

Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj2100717

Published

1983-03-15

Pages

717-20

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1154282

Subset

External Links

PubMed source DOI: 10.1042/bj2100717 PMC Full Text View journal details

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A simple assay method for determination of the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP.

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