Studies on sarcoplasmic reticulum from slow-twitch muscle.
Vesicles were isolated from membranes of the sarcoplasmic reticulum (SR) of rabbit slow-twitch muscle by differential and sucrose density gradient centrifugation after homogenization. In some experiments, the vesicles were further fractionated by loading them with calcium oxalate followed by centrifugation in a sucrose density gradient. Protein composition of the isolated vesicles was complex and differed from the protein composition of fast-twitch muscle vesicles. However, other protein components, which were also present in fast-twitch muscle SR vesicles, have been identified: Ca2 + -dependent ATPase, calsequestrin, 160 000 molecular weight glycoprotein and 53 000 molecular weight glycoprotein. The amount of the Ca2 + -dependent ATPase and calsequestrin was several times lower in the slow-twitch muscle SR vesicles. This has been observed in both the original and the loaded vesicles. The slow-twitch muscle SR vesicles showed active calcium transport, Ca2 + -dependent ATPase activity, and the formation of the phosphorylated intermediate under conditions similar to those established for fast-twitch muscle SR. However, these activities, when expressed per mg of total protein, were several times lower than the analogous activities in the SR vesicles isolated from fast-twitch skeletal muscle. When the same enzyme activities were expressed per mg of the 105 000 molecular weight ATPase, the values obtained were very similar in both kinds of vesicles. The results indicate that the slow rate of calcium transport, found in slow-twitch muscle SR vesicles, may be related to a low content of the calcium-transporting ATPase in the membrane.
D000818:Animals; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002155:Calsequestrin / Q000378:metabolism; D002784:Cholesterol / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis; D007425:Intracellular Membranes / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D009119:Muscle Contraction; D009124:Muscle Proteins / Q000032:analysis; D010743:Phospholipids / Q000032:analysis; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000032:analysis / Q000378:metabolism*; D013997:Time Factors
D002155:Calsequestrin; D006023:Glycoproteins; D008565:Membrane Proteins; D009124:Muscle Proteins; D010743:Phospholipids; D002784:Cholesterol; D000252:Calcium-Transporting ATPases; D002118:Calcium