Am J Physiol Cell Physiol AJP: Heart and Circulatory Physiology
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Am J Physiol Cell Physiol 273: C1915-C1924, 1997;
0363-6143/97 $5.00
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Vol. 273, Issue 6, C1915-C1924, December 1997

Regulation of Ca2+ handling by phosphorylation status in mouse fast- and slow-twitch skeletal muscle fibers

Yewei Liu1, Evangelia G. Kranias2, and Martin F. Schneider1

1 Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201; and 2 Department of Pharmacology and Cell Biophysics, University of Cincinnati, College of Medicine, Cincinnati, Ohio 45267

The effects of phosphorylation status on Ca2+ release and Ca2+ removal were studied in fast-twitch flexor digitorum brevis and slow-twitch soleus skeletal muscle fibers enzymatically isolated from wild-type and phospholamban knockout (PLBko) mice. In all fibers the adenosine 3',5'-cyclic monophosphate-dependent protein kinase (PKA) inhibitor H-89 decreased the peak amplitude of the intracellular Ca2+ concentration ([Ca2+]) transient for a single action potential, and the PKA activator dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) reversed this effect, indicating modulation of Ca2+ release by phosphorylation status in all fibers. H-89 decreased the decay rate constant of the [Ca2+] transient and DBcAMP reversed this effect only in phospholamban-expressing fibers (wild-type soleus), indicating modulation of Ca2+ removal only in the presence of phospholamban. A high basal level of PKA phosphorylation in soleus fibers maintained under our control conditions was indicated by the lack of effect of direct application of DBcAMP on Ca2+ release or Ca2+ removal in wild-type or PLBko soleus fibers and was confirmed by analysis of phospholamban from wild-type soleus fibers.

phospholamban; sarcoplasmic reticulum; calcium release; excitation-contraction coupling


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