Studies on functional domains of the regulatory subunit of bovine heart adenosine 3':5'-monophosphate-dependent protein kinase.

D. A. Flockhart, D. M. Watterson, J. D. Corbin

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Abstract

The functional domains of the regulatory subunit of isozyme II of cAMP-dependent protein kinase were studied. It was shown using Edman degradation that the regulatory subunit contained a phosphorylated residue which was very close in primary sequence to the site most sensitive to hydrolysis by low trypsin concentrations as postulated previously (Corbin, J.D., Sugden, P.H., West, L., Flockhart, D.A., Lincoln, T.M., and McCarthy, D. (1978) J. Biol. Chem. 253, 3997-4003). Catalytic subunit incorporated 0.9 mol of 32P from [gamma-32P]ATP into a preparation of regulatory subunit that contained 1.1 mol of endogenous phosphate. After phosphorylation by the catalytic subunit, the regulatory subunit contained 2.2 mol of chemical phosphate. The effects of heat denaturation upon the rate and extent of phosphorylation of the regulatory subunit were compared with the effects of these treatments upon the cAMP binding and inhibitory domains. These data suggested that the regulatory subunit required factors in addition to an intact phosphorylatable primary sequence in order for inhibitory activity to be expressed. Such factors might be part of the secondary or tertiary structure of the protein. These studies are discussed with respect to the mechanism of inhibition of catalytic activity, and a model of the regulatory subunit structure is proposed.

Original languageEnglish (US)
Pages (from-to)4435-4440
Number of pages6
JournalJournal of Biological Chemistry
Volume255
Issue number10
StatePublished - May 25 1980

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Phosphorylation
Adenosine
Protein Kinases
Catalytic Domain
Phosphates
Denaturation
Cyclic AMP-Dependent Protein Kinases
Tertiary Protein Structure
Trypsin
Isoenzymes
Hydrolysis
Catalyst activity
Hot Temperature
Adenosine Triphosphate
Degradation
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Studies on functional domains of the regulatory subunit of bovine heart adenosine 3':5'-monophosphate-dependent protein kinase. / Flockhart, D. A.; Watterson, D. M.; Corbin, J. D.

In: Journal of Biological Chemistry, Vol. 255, No. 10, 25.05.1980, p. 4435-4440.

Research output: Contribution to journalArticle

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