Cross-linking of the (Ca2+ + Mg2+)-ATPase protein.
The addition of cupric-1,10,-phenanthroline, a cross-linking catalyst, to sarcoplasmic reticulum membranes caused protein sulfhydryl groups to form disulfide bridges. Following a short exposure to the catalyst (15 s, 22 degrees C) most of the protein was in a dimeric form (Mr = 248 000). Longer exposure times resulted in the formation of trimers, tetramers and other oligomers too large to enter the gel. At low temperatures (4 degrees C) dimer formation predominates even for exposure times as long as 5 min. Cross-linking in the presence of 7.5 mM Triton X-100 (a concentration that resulted in clearing of the membrane suspension and thus solubilization of the membrane components) showed the appearance of a considerable dimer fraction, however, most of the (Ca2+ + Mg2+)-ATPase protein appeared as a monomer. Following 1 min of cross-linking at 22 degrees C, freeze-etched membranes showed no alteration in the number or appearance of 80 A intramembranous particles. Thus extensive cross-linking of the (Ca2+ + Mg2+)-ATPase protein can occur without disruption of the normal position of the intramembrane portion of the molecule.
D000818:Animals; D001693:Biological Transport, Active; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D003300:Copper; D004220:Disulfides; D046911:Macromolecular Substances; D008274:Magnesium; D009132:Muscles / Q000378:metabolism; D008121:Nephropidae; D010618:Phenanthrolines; D011092:Polyethylene Glycols; D011485:Protein Binding; D012519:Sarcoplasmic Reticulum / Q000648:ultrastructure
D004220:Disulfides; D046911:Macromolecular Substances; D010618:Phenanthrolines; D011092:Polyethylene Glycols; D003300:Copper; D000252:Calcium-Transporting ATPases; D008274:Magnesium; D002118:Calcium