Specific phosphorylation of a site in the full-length form of the α1 subunit of the cardiac L-type calcium channel by adenosine 3'-5'-cyclic monophosphate-dependent protein kinase

Karen S. De Jongh, Brian J. Murphy, Anita A. Colvin, Johannes W Hell, Masami Takahashi, William A. Catterall

Research output: Contribution to journalArticle

224 Scopus citations

Abstract

Voltage-gated L-type Ca2+ channels mediate Ca2+ entry into cells in response to membrane depolarization Ca2+ entry through the cardiac Ca2+ channel determines the rate and force of contraction, and modulation of Ca2+ channel activity by β-adrenergic agents acting through adenosine 3'- 5'-cyclic monophosphate- (cAMP)-dependent protein phosphorylation contributes to physiological regulation of cardiac function by the sympathetic nervous system. Immunoblotting experiments using the site-directed anti-peptide antibodies against different peptide segments indicate that the α1 subunit of the cardiac L-type Ca2+ channel exists in two size forms with apparent molecular masses of 240 and 210 kDa, which we call α1242 and α1210. α1242 corresponds to the full-length cardiac α1 subunit predicted from its cDNA sequence, while α1210 is truncated at its COOH terminus. Only α1242 is phosphorylated in vitro by cAMP-dependent protein kinase. Protein microsequencing and peptide mapping of wild-type and mutant fusion proteins show that this phosphorylation occurs at serine 1928 near the COOH terminus. Phosphorylation of this residue can be detected by phosphospecific antibodies raised against the corresponding phosphopeptide. Experiments with these antibodies show that α1242 is phosphorylated in intact cells expressing the cardiac α1 subunit in response to increased intracellular levels of cAMP. These results identify serine 1928 on the α1 subunit as a possible site of regulation by cAMP-dependent phosphorylation.

Original languageEnglish (US)
Pages (from-to)10392-10402
Number of pages11
JournalBiochemistry
Volume35
Issue number32
DOIs
StatePublished - 1996
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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