In vivo and in vitro phosphorylation of rat liver fructose-1,6-bisphosphatase

J. P. Riou, T. H. Claus, D. A. Flockhart, J. D. Corbin, S. J. Pilkis

Research output: Contribution to journalArticle

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Abstract

Incorporation of 32P from[γ- 32P] ATP into a homogeneous preparation of rat hepatic fructose-1,6-bisphosphatase (D-fructose-1,6-bisphosphatase 1-phosphohydrolase, EC 3.1.3.11) was catalyzed by a homogeneous preparation of the catalytic subunit of cyclic AMP-dependent protein kinase from bovine liver. Approximately 4 mol of phosphate were incorporated per mol of the tetrameric enzyme. This phosphorylation was associated with an increase in enzyme activity. In addition, in vivo phosphorylation of the enzyme was observed after injection of radioactive inorganic phosphate into rats and subsequent isolation of the enzyme by conventional purification methods and by immunoprecipitation. All of the labeled phosphate incorporation into the enzyme, both in vitro and in vivo, was precipitated by antibody specific for the enzyme. Furthermore, the 32P(i) counts were coincident with the enzyme subunit band when the immunoprecipitates were examined by sodium dodecyl sulfate/disc gel electrophoresis. Acid hydrolysis of the immunoprecipitated enzyme that was phosphorylated in vitro revealed that only seryl residues were labeled. On the basis of the concentration of protein kinase (0.2-1.0 μM) necessary to phosphorylate physiological amounts of fructose-1,6-bisphosphatase (1.0-4.0 μM), it is suggested that cyclic AMP-dependent protein kinase may catalyze the phosphorylation of fructose-1,6-bisphosphatase in vivo.

Original languageEnglish (US)
Pages (from-to)4615-4619
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume74
Issue number10
StatePublished - 1977
Externally publishedYes

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Fructose-Bisphosphatase
Phosphorylation
Liver
Enzymes
Phosphates
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
Disc Electrophoresis
In Vitro Techniques
Cyclic AMP-Dependent Protein Kinases
Phosphoric Monoester Hydrolases
Immunoprecipitation
Sodium Dodecyl Sulfate
Protein Kinases
Hydrolysis
Adenosine Triphosphate
Gels
Injections
Acids
Antibodies

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Riou, J. P., Claus, T. H., Flockhart, D. A., Corbin, J. D., & Pilkis, S. J. (1977). In vivo and in vitro phosphorylation of rat liver fructose-1,6-bisphosphatase. Proceedings of the National Academy of Sciences of the United States of America, 74(10), 4615-4619.

In vivo and in vitro phosphorylation of rat liver fructose-1,6-bisphosphatase. / Riou, J. P.; Claus, T. H.; Flockhart, D. A.; Corbin, J. D.; Pilkis, S. J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 74, No. 10, 1977, p. 4615-4619.

Research output: Contribution to journalArticle

Riou, J. P. ; Claus, T. H. ; Flockhart, D. A. ; Corbin, J. D. ; Pilkis, S. J. / In vivo and in vitro phosphorylation of rat liver fructose-1,6-bisphosphatase. In: Proceedings of the National Academy of Sciences of the United States of America. 1977 ; Vol. 74, No. 10. pp. 4615-4619.
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