Protease digestion of membranes. Ultrastructural and biochemical effects.
(1) Nagarse, a bacterial protease, was permitted to react with sarcoplasmic reticulum, submitochondrial and plasma membranes. Gel electrophoresis indicated that all polypeptides were labile to the enzyme, and therefore must be at least partially exposed at membrane surfaces. However, hydrolysis did not proceed to completion, and in each membrane 30-50% of the original protein mass remained after extensive digestion. Gel patterns showed that remaining polypeptide fragments were in the range of 10000 molecular weight. (2) Amino acid analysis of the original protein and membrane-bound digestion product was performed. Only minor changes were observed following digestion, suggesting that the peptide fragments remaining with the membrane did not have specialized amino acid compositions. (3) freeze-fracture analysis of Nagarse-treated sarcoplasmic and plasma membranes showed that particulate structures were present, although particle density and asymmetry of fistribution between fracture faces were decreased. In submitochondrial membranes, digested membranes were indistinguishable from the original membranes in particle density and distribution. We conclude that high molecular weight polypeptides are not required for the production particulate structures in freeze-fracture images of membranes.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure; D004912:Erythrocytes / Q000648:ultrastructure; D005614:Freeze Fracturing; D006801:Humans; D008566:Membranes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D008861:Microsomes / Q000648:ultrastructure; D008928:Mitochondria / Q000648:ultrastructure; D008121:Nephropidae; D012519:Sarcoplasmic Reticulum / Q000648:ultrastructure; D013381:Subtilisins / Q000494:pharmacology*
D000596:Amino Acids; D013381:Subtilisins