Reversible block of the calcium release channel/ryanodine receptor by protamine, a heparin antidote.

Channel activity of the calcium release channel from skeletal muscle, ryanodine receptor type 1, was measured in the presence and absence of protamine sulfate on the cytoplasmic side of the channel. Single-channel activity was measured after incorporating channels into planar lipid bilayers. Optimally and suboptimally calcium-activated calcium release channels were inactivated by the application of protamine to the cytoplasmic side of the channel. Recovery of channel activity was not observed while protamine was present. The addition of protamine bound to agarose beads did not change channel activity, implying that the mechanism of action involves an interaction with the ryanodine receptor rather than changes in the bulk calcium concentration of the medium. The block of channel activity by protamine could be reversed either by removal by perfusion with buffer or by the addition of heparin to the cytoplasmic side of the channel. Microinjection of protamine into differentiated C(2)C(12) mouse muscle cells prevented caffeine-induced intracellular calcium release. The results suggest that protamine acts on the ryanodine receptor in a similar but opposite manner from heparin and that protamine can be used as a potent, reversible inhibitor of ryanodine receptor activity.

[1]  N. Spitzer,et al.  Calcium signaling in the developing Xenopus myotome. , 1999, Developmental biology.

[2]  R. Mikelsaar,et al.  Are there molecules of nucleoprotamine? , 1999, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  Y. Byun,et al.  Low molecular weight protamine: a potential nontoxic heparin antagonist. , 1999, Thrombosis research.

[4]  J. Mackrill.  Protein-protein interactions in intracellular Ca2+-release channel function , 1999 .

[5]  H. Valdivia,et al.  Modulation of intracellular Ca2+ levels in the heart by sorcin and FKBP12, two accessory proteins of ryanodine receptors. , 1998, TIPS - Trends in Pharmacological Sciences.

[6]  B. Ehrlich,et al.  Characterization of the ryanodine receptor/channel of invertebrate muscle. , 1998, American journal of physiology. Regulatory, integrative and comparative physiology.

[7]  R. Hinojosa,et al.  The systemic vasodilatory action of protamine: is it inhibited or mediated by heparin? , 1997, Research in experimental medicine. Zeitschrift fur die gesamte experimentelle Medizin einschliesslich experimenteller Chirurgie.

[8]  F. Eusebi,et al.  Spontaneous and Repetitive Calcium Transients in C2C12 Mouse Myotubes during In Vitro Myogenesis , 1997, The European journal of neuroscience.

[9]  H. Wässle,et al.  Group I Metabotropic Glutamate Receptors mGluR1α and mGluR5a: Localization in Both Synaptic Layers of the Rat Retina , 1997, The Journal of Neuroscience.

[10]  M. Berridge,et al.  Expression and Function of Ryanodine Receptors in Nonexcitable Cells (*) , 1996, The Journal of Biological Chemistry.

[11]  D. Maclennan,et al.  Identification of calmodulin-, Ca(2+)-, and ruthenium red-binding domains in the Ca2+ release channel (ryanodine receptor) of rabbit skeletal muscle sarcoplasmic reticulum. , 1994, The Journal of biological chemistry.

[12]  M. Fill,et al.  Surface charge potentiates conduction through the cardiac ryanodine receptor channel , 1994, The Journal of general physiology.

[13]  I. Bezprozvanny,et al.  The pharmacology of intracellular Ca2+-release channels , 1994 .

[14]  I. Bezprozvanny,et al.  Intracellular calcium release channels. , 1994, The Chinese journal of physiology.

[15]  S. Baylor,et al.  Resting myoplasmic free calcium in frog skeletal muscle fibers estimated with fluo-3. , 1993, Biophysical journal.

[16]  D. Maclennan,et al.  Antibodies as probes for Ca2+ activation sites in the Ca2+ release channel (ryanodine receptor) of rabbit skeletal muscle sarcoplasmic reticulum. , 1993, The Journal of biological chemistry.

[17]  I. Bezprozvanny,et al.  Activation of the calcium release channel (ryanodine receptor) by heparin and other polyanions is calcium dependent. , 1993, Molecular biology of the cell.

[18]  D. Maclennan,et al.  Characterization of a Ca2+ binding and regulatory site in the Ca2+ release channel (ryanodine receptor) of rabbit skeletal muscle sarcoplasmic reticulum. , 1992, The Journal of biological chemistry.

[19]  T. Itoh,et al.  Effects of protamine on vascular smooth muscle of rabbit mesenteric artery. , 1991, Anesthesiology.

[20]  James Watras,et al.  Bell-shaped calcium-response curves of lns(l,4,5)P3- and calcium-gated channels from endoplasmic reticulum of cerebellum , 1991, Nature.

[21]  T. Südhof,et al.  Putative receptor for inositol 1,4,5-trisphosphate similar to ryanodine receptor , 1989, Nature.

[22]  P. Tempst,et al.  Molecular cloning and characterization of the ryanodine receptor/junctional channel complex cDNA from skeletal muscle sarcoplasmic reticulum. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[23]  P. Palade,et al.  Pharmacologic differentiation between inositol-1,4,5-trisphosphate-induced Ca2+ release and Ca2+- or caffeine-induced Ca2+ release from intracellular membrane systems. , 1989, Molecular pharmacology.

[24]  H. Takeshima,et al.  Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor , 1989, Nature.

[25]  J. Watras,et al.  Inositol 1,4,5-trisphosphate activates a channel from smooth muscle sarcoplasmic reticulum , 1988, Nature.

[26]  J. Mullaney,et al.  Competitive, reversible, and potent antagonism of inositol 1,4,5-trisphosphate-activated calcium release by heparin. , 1988, Journal of Biological Chemistry.

[27]  S H Snyder,et al.  Solubilization, purification, and characterization of an inositol trisphosphate receptor. , 1988, The Journal of biological chemistry.

[28]  P. Palade.  Drug-induced Ca2+ release from isolated sarcoplasmic reticulum. III. Block of Ca2+-induced Ca2+ release by organic polyamines. , 1987, The Journal of biological chemistry.

[29]  G. Meissner,et al.  Single channel measurements of the calcium release channel from skeletal muscle sarcoplasmic reticulum. Activation by Ca2+ and ATP and modulation by Mg2+ , 1986, The Journal of general physiology.

[30]  N. Ikemoto,et al.  Kinetic studies of calcium release from sarcoplasmic reticulum in vitro. , 1983, The Journal of biological chemistry.

[31]  K. Suzuki,et al.  Studies on protamines. XVI. The complete amino acid sequence of clupeine YII. , 1972, Journal of biochemistry.

[32]  K. Iwai,et al.  Studies on protamines. XV. The complete amino acid sequence of the Z component of clupeine. Application of N leads to O acyl rearrangement and selective hydrolysis in sequence determination. , 1971, Journal of biochemistry.

[33]  M. Yamasaki,et al.  STUDIES ON PROTAMINES , 1958 .

[34]  J. Mackrill.  Protein-protein interactions in intracellular Ca2+-release channel function. , 1999, The Biochemical journal.

[35]  I. Bezprozvanny,et al.  The pharmacology of intracellular Ca(2+)-release channels. , 1994, Trends in pharmacological sciences.

[36]  A. Fabiato,et al.  Computer programs for calculating total from specified free or free from specified total ionic concentrations in aqueous solutions containing multiple metals and ligands. , 1988, Methods in enzymology.

[37]  P. Palade.  Drug-induced Ca'' Release from Isolated Sarcoplasmic Reticulum , 1987 .

[38]  K. Suzuki,et al.  Studies on protamines. XVII. The complete amino acid sequence of clupeine YI. , 1972, Journal of biochemistry.