Calcium-ryanodine receptor complex: solubilization and partial characterization

Conference · Fri Feb 28 23:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5385266

Pessah, I N; Francini, A O; Casida, J E

The Ca/sup 2 +/-ryanodine receptor complex is solubilized on treating sarcoplasmic reticulum (SR) vesicles from skeletal muscles with 3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate and NaCl. The heavy membrane preparations obtained from pyrophosphate homogenates frequently exhibit junctional feet and are derived from the ternal cisternae of the SR. (/sup 3/H)Ryanodine binding to the soluble receptor and the heavy SR vesicles is similar with respect to dependent on Ca/sup 2 +/, inhibition by Mg/sup 2 +/, pharmacological specificity for inhibition by six ryanoids and ruthenium red, and lack of sensitivity to voltage-dependent Ca/sup 2 +/-channel blockers, inositol 1,4,5-trisphosphate, or doxorubicin. In contrast, the cation sensitivity is decreased on solubilization of the skeletal receptor to a level more characteristic of the membrane-associated cardiac receptor. The soluble skeletal receptor is modulated by nucleotides and rapidly denatured at 50/sup 0/C. Binding is characterized by association and dissociation constants of 0.028 min/sup -1/ and 0.034 hr/sup -1/, respectively. Polyacrylamide gel electrophoresis and Sepharose chromatography show that the receptor complex consists primarily of two protein fractions, one of apparent M/sub r/ 150,000-300,000, and a second, the (/sup 3/H)ryanodine binding component of M/sub r/ 1.2 x 10/sup 6/. Proteins evident on analysis by SDS-PAGE indicate that the former fraction is enriched in calsequestrin (M/sub r/ 64,000) and the latter is made up of units of M/sub r/ 104,000, 227,000 and 438,000.

• Research Organization: Univ. of California, Berkeley
• OSTI ID: 5385266
• Report Number(s): CONF-8604222-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:3; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METALS
BIOCHEMICAL REACTION KINETICS
BODY
CALCIUM
CARDIOVASCULAR SYSTEM
CATIONS
CELL CONSTITUENTS
CELL MEMBRANES
CHARGED PARTICLES
CHROMATOGRAPHY
ELECTROPHORESIS
ELEMENTS
HEART
INSECTICIDES
IONS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAGNESIUM
MEMBRANE PROTEINS
MEMBRANES
METALS
MUSCLES
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
OXYGEN COMPOUNDS
PESTICIDES
PHOSPHORUS COMPOUNDS
PROTEINS
PYROPHOSPHATES
REACTION KINETICS
RECEPTORS
SARCOPLASMIC RETICULUM
SEPARATION PROCESSES
SKELETON
TRACER TECHNIQUES
TRITIUM COMPOUNDS