Altered properties of calsequestrin and the ryanodine receptor in the cardiac sarcoplasmic reticulum of hibernating mammals.

A novel isoform of calsequestrin was identified in sarcoplasmic reticulum vesicles from myocardial tissue of two species of hibernating ground squirrel. The protein was identified as calsequestrin by its cross-reactivity with antibodies raised against bovine cardiac calsequestrin, its pH-sensitive mobility in sodium dodecylsulphate-polyacrylamide gels, staining blue with the cationic carbocyanine dye 'Stains-All', binding peroxidase-conjugated concanavalin A, its endoglycosidase F sensitivity. Its NH2-terminal amino acid sequence is similar, but not identical, to that already determined for cardiac calsequestrin. Some of the biochemical properties of this protein distinguish it from the other mammalian isoforms. It has a unique electrophoretic mobility in both alkaline and neutral sodium dodecylsulphate-polyacrylamide gel electrophoresis, it appears to have a molecular weight approximately 7% greater than that of cardiac calsequestrin from other mammalian species, and its glycosylation pattern differs. This novel form of calsequestrin is expressed in cardiac SR vesicles which possess an abnormally high number of Ca2(+)-release channel/ryanodine receptor molecules. This ryanodine receptor also shows an altered Ca2(+)-sensitivity of ryanodine binding. The divergent biophysical properties of this novel form of cardiac calsequestrin, together with the apparently atypical ryanodine receptors in the cardiac sarcoplasmic reticulum membranes may have some functional significance in the adaptive mechanisms which allow the heart to function despite the severely reduced body temperatures (to approx. 0 degree C) encountered during hibernation.