PMID: 3768353Oct 23, 1986Paper

Ca2+-dependent ryanodine binding site: soluble preparation from rabbit cardiac sarcoplasmic reticulum

Biochimica Et Biophysica Acta
J Seifert, J E Casida

Abstract

The Ca2+-dependent ryanodine binding site of rabbit cardiac sarcoplasmic reticulum is solubilized by treatment with 20 mM CHAPS detergent and 1 M NaCl for 30 min at 0 degrees C. Ca2+ added at 5 microM enhances binding, at 0.5 mM increases both the affinity and number of [3H]ryanodine binding sites, while at 10 mM only the number of binding sites is increased. Mg2+ up to 1 mM does not significantly affect [3H]ryanodine binding. Radioligand binding is strongly enhanced by all alkali metal chlorides except LiCl. NaCl increases the rate of association of the ligand and the affinity of the binding site but does not influence the dissociation. NaCl and CaCl2 enhance the thermal stability of the [3H]ryanodine-binding protein. Thiol groups are essential for [3H]ryanodine binding. Ruthenium red and Cd2+ inhibit binding, while theophylline is stimulatory at low (micromolar) Ca2+ concentrations by a mechanism other than phosphodiesterase inhibition. Gel permeation chromatography establishes that the ryanodine binding protein is localized only in the high molecular mass fraction (greater than 669 kDa). Polyacrylamide gel electrophoresis of the proteins following treatment with SDS and 2-mercaptoethanol indicates that more than 90% are of l...Continue Reading

References

Feb 28, 1985·Biochimica Et Biophysica Acta·J J Feher, G B Lipford
Apr 16, 1985·Biochemical and Biophysical Research Communications·I N PessahJ E Casida
Mar 11, 1974·Pflügers Archiv : European journal of physiology·Z J Penefsky
Mar 1, 1971·Journal of Medicinal Chemistry·A R PatelR E Lutz
Apr 1, 1983·The Journal of Cell Biology·R D MitchellS Fleischer
Jul 1, 1983·Analytical Biochemistry·R F BrunsT A Pugsley

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Citations

Apr 1, 1989·Journal of Bioenergetics and Biomembranes·F A Lai, G Meissner
Feb 1, 1993·The Journal of Membrane Biology·N R BrandtA L Bassett
Jun 26, 2007·Acta Pharmacologica Sinica·Marco Santonastasi, Xander H T Wehrens
Oct 12, 2010·Seminars in Cell & Developmental Biology·Uwe SchulteBernd Fakler
Apr 21, 2019·The Journal of General Physiology·Maedeh Bazmi, Ariel L Escobar
Feb 1, 1992·Journal of Muscle Research and Cell Motility·A J Williams
Apr 22, 1988·Biochimica Et Biophysica Acta·M MichalakV Shoshan-Barmatz

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