PMID: 6288733Jun 1, 1982

Direct cytochemical localization of catalytic subunits dissociated from cAMP-dependent protein kinase in Reuber H-35 hepatoma cells. II. Temporal and spatial kinetics

The Journal of Cell Biology
C V Byus, W H Fletcher

Abstract

The activation of cyclic AMP-dependent protein kinase has been found to be the predominant mode by which cyclic AMP (cAMP) leads to alterations of a large variety of cellular functions. The activation of the kinase results in the release of the catalytic subunit which as the free enzyme possesses phosphotransferase activity for a variety of specific protein substrates. Using a sensitive and specific cytofluorometric technique we monitored the appearance of free catalytic subunit in Reuber H35 hepatoma cells in culture after incubation with N6-1'-O-dibutyryl-cyclic AMP (DBcAMP), 8-bromoadenosine-3':5'-cyclic monophosphate (8-BrcAMP), and glucagon. The cytochemical method employs the heat-stable inhibitor of the free catalytic subunit which has been conjugated to fluorescein isothiocyanate (F:PKI) and was validated as described in the companion paper (Fletcher and Byus. 1982. J. Cell Biol. 93:719-726). Here we studied the temporal and spatial kinetics of the free catalytic subunit following activation of cAMP-dependent protein kinase by increasing concentrations of DBcAMP,8-BrcAMP, and glucagon. Under similar conditions protein kinase activation was also assessed biochemically in H35 cell supernatants by assaying the protein kina...Continue Reading

References

Apr 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·E CostaA Guidotti
Apr 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·S L KeelyC R Park
Dec 6, 1976·Biochemical and Biophysical Research Communications·C V ByusD H Russell
Jan 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·D J FullerD H Russell
Jun 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·M K HaddoxD H Russell
Jan 1, 1980·Annual Review of Pharmacology and Toxicology·D B Glass, E G Krebs

Citations

Nov 1, 1993·Molecular and Cellular Biochemistry·J L MeinkothJ R Feramisco
Jul 1, 1985·Molecular and Cellular Biochemistry·G A ClawsonE A Smuckler
Sep 1, 1986·Molecular and Cellular Endocrinology·S A Murray, W H Fletcher
Nov 1, 1995·Critical Reviews in Oncology/hematology·Y S Cho-ChungM Nesterova
Dec 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·J L MeinkothJ R Feramisco
May 1, 1984·The Journal of Cell Biology·S A Murray, W H Fletcher
Feb 11, 2004·The Journal of Cell Biology·Jonathan H ZippinJochen Buck
Jan 23, 2002·Developmental Dynamics : an Official Publication of the American Association of Anatomists·John C Duncan, W H Fletcher

Related Concepts

CGKIbeta
8-bromo-2'-AMP
Cyclic AMP, (R)-Isomer
Phosphaden
Carrier Proteins
Catalysis
Cell Nucleolus
Protoplasm
Bucladesine, Barium (1: 1) Salt
Enzyme Activation

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