Literature DB >> 6307269

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

P T Hawkins, R H Michell, C J Kirk.   

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.

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Year:  1983        PMID： 6307269      PMCID： PMC1154282          DOI： 10.1042/bj2100717

Source DB:  PubMed          Journal:  Biochem J        ISSN： 0264-6021            Impact factor:   3.857


  6 in total

1.  PURIFICATION AND PROPERTIES OF RABBIT SKELETAL MUSCLE PHOSPHORYLASE B KINASE.

Authors:  E G KREBS; D S LOVE; G E BRATVOLD; K A TRAYSER; W L MEYER; E H FISCHER
Journal:  Biochemistry       Date:  1964-08       Impact factor: 3.162

2.  Assay of cyclic AMP-dependent protein kinases.

Authors:  J D Corbin; E M Reimann
Journal:  Methods Enzymol       Date:  1974       Impact factor: 1.600

3.  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.

Authors:  J C Garrison; M K Borland; V A Florio; D A Twible
Journal:  J Biol Chem       Date:  1979-08-10       Impact factor: 5.157

4.  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.

Authors:  P J England; D A Walsh
Journal:  Anal Biochem       Date:  1976-10       Impact factor: 3.365

5.  Phosphatidylinositol metabolism in rat hepatocytes stimulated by vasopressin.

Authors:  C J Kirk; R H Michell; D A Hems
Journal:  Biochem J       Date:  1981-01-15       Impact factor: 3.857

6.  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.

Authors:  J M Cogoli; J G Dobson
Journal:  Anal Biochem       Date:  1981-01-15       Impact factor: 3.365
  6 in total
  13 in total

1.  Multiple metabolic pools of phosphoinositides and phosphatidate in human erythrocytes incubated in a medium that permits rapid transmembrane exchange of phosphate.

Authors:  C E King; L R Stephens; P T Hawkins; G R Guy; R H Michell
Journal:  Biochem J       Date:  1987-05-15       Impact factor: 3.857

2.  Experimental determination of the phosphorylation state of phenylalanine hydroxylase.

Authors:  A K Green; R G Cotton; I Jennings; M J Fisher
Journal:  Biochem J       Date:  1990-01-15       Impact factor: 3.857

3.  The role of insulin in the modulation of glucagon-dependent control of phenylalanine hydroxylation in isolated liver cells.

Authors:  M J Fisher; A J Dickson; C I Pogson
Journal:  Biochem J       Date:  1987-03-15       Impact factor: 3.857

4.  The labelling of polyphosphoinositides with [32P]Pi and the accumulation of inositol phosphates in vasopressin-stimulated hepatocytes.

Authors:  S Palmer; P T Hawkins; R H Michell; C J Kirk
Journal:  Biochem J       Date:  1986-09-01       Impact factor: 3.857

5.  An analysis of myo-[3H]inositol trisphosphates found in myo-[3H]inositol prelabelled avian erythrocytes.

Authors:  L R Stephens; P T Hawkins; C P Downes
Journal:  Biochem J       Date:  1989-09-15       Impact factor: 3.857

6.  Effects of adrenergic agents, vasopressin and ionophore A23187, on the phosphorylation of, and flux through, phenylalanine hydroxylase in rat liver cells.

Authors:  M J Fisher; M A Santana; C I Pogson
Journal:  Biochem J       Date:  1984-04-01       Impact factor: 3.857

7.  Impaired nitric oxide-dependent cyclic guanosine monophosphate generation in glomeruli from diabetic rats. Evidence for protein kinase C-mediated suppression of the cholinergic response.

Authors:  P A Craven; R K Studer; F R DeRubertis
Journal:  J Clin Invest       Date:  1994-01       Impact factor: 14.808

8.  Phenylalanine hydroxylase in liver cells. Correlation of glucagon-stimulated enzyme phosphorylation with expressed activity.

Authors:  M J Fisher; C I Pogson
Journal:  Biochem J       Date:  1984-04-01       Impact factor: 3.857

9.  Okadaic acid identifies a phosphorylation/dephosphorylation cycle controlling the inhibitory guanine-nucleotide-binding regulatory protein Gi2.

Authors:  M Bushfield; B E Lavan; M D Houslay
Journal:  Biochem J       Date:  1991-03-01       Impact factor: 3.857

10.  The effect of tetrahydrobiopterin on the in situ phosphorylation of tyrosine hydroxylase in rat striatal synaptosomes.

Authors:  P Ribeiro; S Kaufman
Journal:  Neurochem Res       Date:  1994-05       Impact factor: 3.996
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