β-Adrenergic stimulation of phospholamban phosphorylation and Ca 2+-ATPase activity in guinea pig ventricles

J. P. Lindemann, Larry Jones, D. R. Hathaway, B. G. Henry, A. M. Watanabe

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Abstract

The effects of β-adrenergic stimulation on phospholamban phosphorylation and Ca 2+-ATPase activity were studied in intact myocarduim. Isolated guinea pig ventricles were perfused via the coronary arteries with 32P(i) after which membrane vesicles, comprised primarily of sarcoplasmic reticulum, were isolated by differential centrifugation. Isoproterenol perfusion increased 32P incorporation into membrane proteins of apparent M(r)=22.000 and 8,000, which have been shown to be the predominant proteins phosphorylated in sarcoplasmic reticulum by cAMP-dependent protein kinase. 32P incorporation was increased 3.8-fold and 7.1-fold after 60-s exposure to 10 and 100 nM isoproterenol, respectively. In the same membrane vesicles, Ca 2+-ATPase activity was increased by 30 and 52%, respectively. The increases in phospholamban phosphorylation and Ca 2+-ATPase activity correlated in time and concentration dependence with increases in the rate of myocardial relaxation. These biochemical changes did not correlate well with positive inotropy. Similar effects were produced by histamine, 1μM. The onset of the effects of isoproterenol on phospholamban phosphorylation and Ca 2+-ATPase activity closely paralleled the relaxation effects. However, reversal of the relaxant effects occurred more rapidly than the return of 32P incorporation and Ca 2+-ATPase activity to control levels. These results provide evidence that β-adrenergic stimulation results in phospholamban phosphorylation and increased Ca 2+-ATPase activity in intact, functioning ventricular myocardium. These findings support the hypothesis that stimulation of sarcoplasmic reticulum Ca 2+ transport, mediated by cAMP-dependent phosphorylation of phospolamban, may underlie the relaxant effects of β-adrenergic stimulation in mammalian myocardium.

Original languageEnglish
Pages (from-to)464-471
Number of pages8
JournalJournal of Biological Chemistry
Volume258
Issue number1
StatePublished - 1983
Externally publishedYes

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Phosphorylation
Adrenergic Agents
Adenosine Triphosphatases
Guinea Pigs
Sarcoplasmic Reticulum
Isoproterenol
Myocardium
Membranes
Centrifugation
Level control
Cyclic AMP-Dependent Protein Kinases
Histamine
phospholamban
Coronary Vessels
Membrane Proteins
Perfusion
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

β-Adrenergic stimulation of phospholamban phosphorylation and Ca 2+-ATPase activity in guinea pig ventricles. / Lindemann, J. P.; Jones, Larry; Hathaway, D. R.; Henry, B. G.; Watanabe, A. M.

In: Journal of Biological Chemistry, Vol. 258, No. 1, 1983, p. 464-471.

Research output: Contribution to journalArticle

Lindemann, J. P. ; Jones, Larry ; Hathaway, D. R. ; Henry, B. G. ; Watanabe, A. M. / β-Adrenergic stimulation of phospholamban phosphorylation and Ca 2+-ATPase activity in guinea pig ventricles. In: Journal of Biological Chemistry. 1983 ; Vol. 258, No. 1. pp. 464-471.
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