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Interaction of ryanodine with canine cardiac sarcoplasmic reticulum

Thesis/Dissertation ·
OSTI ID:7027117
Ryanodine was used as a tool to study the Ca release channel of canine cardiac sarcoplasmic reticulum. The binding of /sup 3/H-ryanodine to cardiac sarcoplasmic reticulum vesicles at 37/sup 0/C was investigated by Millipore filtration and liquid scintillation spectrometry. The binding of /sup 3/H-ryanodine depended on the presence of Ca. The Hill coefficient for the Ca requirement for /sup 3/H-ryanodine binding averaged 2.09, indicating that a minimum of two Ca ions were required to bind to specific Ca receptors prior to the binding of ryanodine, and that the binding of the Ca ions was positively cooperative. The K/sub m/ for Ca decreased from 1.97 +/- 0.30 to 0.77 +/- 0.03 nM when the ryanodine concentration was increased from 11 to 268 nM. Scatchard analysis indicated the presence of a single high-affinity binding site having an average K/sub d/ of 6.95 +/- 0.44 nM ryanodine and an average B/sub max/ of 5.34 +/- 0.34 pmol/mg. The analysis also suggested the existence of a second, low-affinity site. Ryanodine binding to the low-affinity site was estimated from the effect of ryanodine on the oxalate-supported Ca uptake rate measured by Millipore filtration with /sup 45/Ca tracer.
Research Organization:
Virginia Commonwealth Univ., Richmond (USA)
OSTI ID:
7027117
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH ISOTOPES
ALKALINE EARTH METALS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CALCIUM
CALCIUM 45
CALCIUM ISOTOPES
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
DOGS
ELEMENTS
EVEN-ODD NUCLEI
HEART
INSECTICIDES
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
METALS
NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
PESTICIDES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
SARCOPLASMIC RETICULUM
TRACER TECHNIQUES
TRITIUM COMPOUNDS
UPTAKE
VERTEBRATES