Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1991 Oct;60(4):902–909. doi: 10.1016/S0006-3495(91)82124-5

Dihydropyridine-sensitive skeletal muscle Ca channels in polarized planar bilayers. 2. Effects of phosphorylation by cAMP-dependent protein kinase.

C Mundiña-Weilenmann 1, J Ma 1, E Ríos 1, M M Hosey 1
PMCID: PMC1260141  PMID: 1660320

Abstract

The effects of phosphorylation on the voltage-dependent properties of dihydropyridine-sensitive Ca channels of skeletal muscle were studied. Single channel currents were recorded upon incorporation of transverse tubule membranes into planar bilayers that were kept polarized at near physiological resting potential and subjected to depolarizing pulses under voltage clamp. Studies were conducted to analyze the properties of the channels at both the single channel and macroscopic level, using methods introduced in the preceding paper (Ma et al., 1991. Biophys. J. 60: 890-901.). Addition of the catalytic subunit of cAMP-dependent protein kinase to the cis (intracellular) side of the bilayers containing channels resulted in: (a) an increase in open channel probability at all voltages above -50 mV; (b) a leftward shift (by 7 mV) in the curve describing the voltage-dependence of activation; (c) an approximate twofold decrease in the rate of inactivation; and (d) an increase in the availability of the channel. These findings provide new insights at the single channel level into the mechanism of modulation of the dihydropyridine-sensitive Ca channels of skeletal muscle by signal transduction events that involve elevation in cAMP and activation of the cAMP-dependent protein kinase.

Full text

PDF
903

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arreola J., Calvo J., García M. C., Sánchez J. A. Modulation of calcium channels of twitch skeletal muscle fibres of the frog by adrenaline and cyclic adenosine monophosphate. J Physiol. 1987 Dec;393:307–330. doi: 10.1113/jphysiol.1987.sp016825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown A. M., Birnbaumer L. Direct G protein gating of ion channels. Am J Physiol. 1988 Mar;254(3 Pt 2):H401–H410. doi: 10.1152/ajpheart.1988.254.3.H401. [DOI] [PubMed] [Google Scholar]
  3. Cleland W. W. The statistical analysis of enzyme kinetic data. Adv Enzymol Relat Areas Mol Biol. 1967;29:1–32. doi: 10.1002/9780470122747.ch1. [DOI] [PubMed] [Google Scholar]
  4. Curtis B. M., Catterall W. A. Reconstitution of the voltage-sensitive calcium channel purified from skeletal muscle transverse tubules. Biochemistry. 1986 Jun 3;25(11):3077–3083. doi: 10.1021/bi00359a002. [DOI] [PubMed] [Google Scholar]
  5. Di Virgilio F., Salviati G., Pozzan T., Volpe P. Is a guanine nucleotide-binding protein involved in excitation-contraction coupling in skeletal muscle? EMBO J. 1986 Feb;5(2):259–262. doi: 10.1002/j.1460-2075.1986.tb04207.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Flockerzi V., Oeken H. J., Hofmann F., Pelzer D., Cavalié A., Trautwein W. Purified dihydropyridine-binding site from skeletal muscle t-tubules is a functional calcium channel. Nature. 1986 Sep 4;323(6083):66–68. doi: 10.1038/323066a0. [DOI] [PubMed] [Google Scholar]
  7. García J., Gamboa-Aldeco R., Stefani E. Charge movement and calcium currents in skeletal muscle fibers are enhanced by GTP gamma S. Pflugers Arch. 1990 Sep;417(1):114–116. doi: 10.1007/BF00370779. [DOI] [PubMed] [Google Scholar]
  8. Gonzalez-Serratos H., Hill L., Valle-Aguilera R. Effects of catecholamines and cyclic amp on excitation--contraction coupling in isolated skeletal muscle fibres of the frog. J Physiol. 1981 Jun;315:267–282. doi: 10.1113/jphysiol.1981.sp013747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hosey M. M., Lazdunski M. Calcium channels: molecular pharmacology, structure and regulation. J Membr Biol. 1988 Sep;104(2):81–105. doi: 10.1007/BF01870922. [DOI] [PubMed] [Google Scholar]
  10. Jahn H., Nastainczyk W., Röhrkasten A., Schneider T., Hofmann F. Site-specific phosphorylation of the purified receptor for calcium-channel blockers by cAMP- and cGMP-dependent protein kinases, protein kinase C, calmodulin-dependent protein kinase II and casein kinase II. Eur J Biochem. 1988 Dec 15;178(2):535–542. doi: 10.1111/j.1432-1033.1988.tb14480.x. [DOI] [PubMed] [Google Scholar]
  11. Lai Y., Seagar M. J., Takahashi M., Catterall W. A. Cyclic AMP-dependent phosphorylation of two size forms of alpha 1 subunits of L-type calcium channels in rat skeletal muscle cells. J Biol Chem. 1990 Dec 5;265(34):20839–20848. [PubMed] [Google Scholar]
  12. Mundiña-Weilenmann C., Chang C. F., Gutierrez L. M., Hosey M. M. Demonstration of the phosphorylation of dihydropyridine-sensitive calcium channels in chick skeletal muscle and the resultant activation of the channels after reconstitution. J Biol Chem. 1991 Mar 5;266(7):4067–4073. [PubMed] [Google Scholar]
  13. Nastainczyk W., Röhrkasten A., Sieber M., Rudolph C., Schächtele C., Marmè D., Hofmann F. Phosphorylation of the purified receptor for calcium channel blockers by cAMP kinase and protein kinase C. Eur J Biochem. 1987 Nov 16;169(1):137–142. doi: 10.1111/j.1432-1033.1987.tb13590.x. [DOI] [PubMed] [Google Scholar]
  14. Nunoki K., Florio V., Catterall W. A. Activation of purified calcium channels by stoichiometric protein phosphorylation. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6816–6820. doi: 10.1073/pnas.86.17.6816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. O'Callahan C. M., Hosey M. M. Multiple phosphorylation sites in the 165-kilodalton peptide associated with dihydropyridine-sensitive calcium channels. Biochemistry. 1988 Aug 9;27(16):6071–6077. doi: 10.1021/bi00416a036. [DOI] [PubMed] [Google Scholar]
  16. O'Callahan C. M., Ptasienski J., Hosey M. M. Phosphorylation of the 165-kDa dihydropyridine/phenylalkylamine receptor from skeletal muscle by protein kinase C. J Biol Chem. 1988 Nov 25;263(33):17342–17349. [PubMed] [Google Scholar]
  17. Ochi R., Kawashima Y. Modulation of slow gating process of calcium channels by isoprenaline in guinea-pig ventricular cells. J Physiol. 1990 May;424:187–204. doi: 10.1113/jphysiol.1990.sp018062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pelzer D., Pelzer S., McDonald T. F. Properties and regulation of calcium channels in muscle cells. Rev Physiol Biochem Pharmacol. 1990;114:107–207. doi: 10.1007/BFb0031019. [DOI] [PubMed] [Google Scholar]
  19. Rios E., Brum G. Involvement of dihydropyridine receptors in excitation-contraction coupling in skeletal muscle. Nature. 1987 Feb 19;325(6106):717–720. doi: 10.1038/325717a0. [DOI] [PubMed] [Google Scholar]
  20. Röhrkasten A., Meyer H. E., Nastainczyk W., Sieber M., Hofmann F. cAMP-dependent protein kinase rapidly phosphorylates serine- 687 of the skeletal muscle receptor for calcium channel blockers. J Biol Chem. 1988 Oct 25;263(30):15325–15329. [PubMed] [Google Scholar]
  21. Schmid A., Renaud J. F., Lazdunski M. Short term and long term effects of beta-adrenergic effectors and cyclic AMP on nitrendipine-sensitive voltage-dependent Ca2+ channels of skeletal muscle. J Biol Chem. 1985 Oct 25;260(24):13041–13046. [PubMed] [Google Scholar]
  22. Sugden P. H., Holladay L. A., Reimann E. M., Corbin J. D. Purification and characterization of the catalytic subunit of adenosine 3':5'-cyclic monophosphate-dependent protein kinase from bovine liver. Biochem J. 1976 Nov;159(2):409–422. doi: 10.1042/bj1590409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tanabe T., Beam K. G., Adams B. A., Niidome T., Numa S. Regions of the skeletal muscle dihydropyridine receptor critical for excitation-contraction coupling. Nature. 1990 Aug 9;346(6284):567–569. doi: 10.1038/346567a0. [DOI] [PubMed] [Google Scholar]
  24. Tanabe T., Beam K. G., Powell J. A., Numa S. Restoration of excitation-contraction coupling and slow calcium current in dysgenic muscle by dihydropyridine receptor complementary DNA. Nature. 1988 Nov 10;336(6195):134–139. doi: 10.1038/336134a0. [DOI] [PubMed] [Google Scholar]
  25. Tanabe T., Takeshima H., Mikami A., Flockerzi V., Takahashi H., Kangawa K., Kojima M., Matsuo H., Hirose T., Numa S. Primary structure of the receptor for calcium channel blockers from skeletal muscle. Nature. 1987 Jul 23;328(6128):313–318. doi: 10.1038/328313a0. [DOI] [PubMed] [Google Scholar]
  26. Tsien R. W., Bean B. P., Hess P., Lansman J. B., Nilius B., Nowycky M. C. Mechanisms of calcium channel modulation by beta-adrenergic agents and dihydropyridine calcium agonists. J Mol Cell Cardiol. 1986 Jul;18(7):691–710. doi: 10.1016/s0022-2828(86)80941-5. [DOI] [PubMed] [Google Scholar]
  27. Yatani A., Imoto Y., Codina J., Hamilton S. L., Brown A. M., Birnbaumer L. The stimulatory G protein of adenylyl cyclase, Gs, also stimulates dihydropyridine-sensitive Ca2+ channels. Evidence for direct regulation independent of phosphorylation by cAMP-dependent protein kinase or stimulation by a dihydropyridine agonist. J Biol Chem. 1988 Jul 15;263(20):9887–9895. [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES