Studies on the heterogeneity of sarcoplasmic reticulum vesicles.
Isolated sarcoplasmic reticulum vesicles from rabbit white muscle were separated into a light (15--20% of total microsomes) and a heavy (80--85%) fraction by density gradient centifugation. The ultrastructure, chemical composition, enzymic activities and localization of membrane components in the vesicles of both fractions were investigated. From the following results it was concluded that both fractions are derived from the membranes of the sarcoplasmic reticulum system of the muscle: (i) The protein pattern of both fractions is essentially the same, except for different ratios of acidic, Ca2+-binding proteins. (ii) The 105000 dalton protein of the light fraction cross-reacts immunologically with the Ca2+-dependent ATPase of the heavy fraction. (iii) Ca2+-dependent ATPase, although of different specific activity, is found in both fractions. After rendering the vesicles leaky, specific activities in both fractions reach the same value. The light fraction was found to consist of "inside-out" vesicles by the following criteria: (i) No Ca2+ accumulation can be measured and the Ca2+-dependent ATPase activity is low and variable. (ii) The rate of trypsin digestion is lower and, compared to the heavy microsomes, a different ratio of degradation products is obtained. (iii) The sarcoplasmic reticulum membrane has a highly asymmetrical lipid distribution. This distribution of aminophospholipids is opposite to that in vesicles of heavy fraction. The light sarcoplasmic reticulum fraction has a higher phospholipid to protein ratio than the heavy one. This is consistent with the possibility that the two fractions derive from different parts of the sarcoplasmic reticulum system.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002474:Cell-Free System; D008055:Lipids / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D009124:Muscle Proteins / Q000032:analysis; D009132:Muscles / Q000648:ultrastructure; D010740:Phospholipases / Q000378:metabolism; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure*; D014302:Trinitrobenzenesulfonic Acid
D008055:Lipids; D008565:Membrane Proteins; D009124:Muscle Proteins; D014302:Trinitrobenzenesulfonic Acid; D010740:Phospholipases; D000251:Adenosine Triphosphatases; D002118:Calcium