Immunochemical analysis of triton X-100-insoluble residues from Micrococcus lysodeikticus membranes.
Triton X-100-insoluble residues from Micrococcus lysodeikticus membranes were analyzed by crossed immunoelectrophoresis after dispersal of the residues in sodium dodecyl sulfate (SDS). Conditions which produce no obvious distortion of the immunoprecipitate profile and which allow qualitative and quantitative analyses of the antigens present in the extracts are described. Two main antigens were detected; these were identified as succinate dehydrogenase (EC 1.3.99.1) and adenosine triphosphatase (EC 3.6.1.3). As determined by peak area estimations, the maximal release of succinate dehydrogenase and of adenosine triphosphatase from Triton X-100-insoluble membrane residues occurred at protein/SDS ratios of about 4.3:1 (0.2% SDS) and 6.8:1 (0.13% SDS), respectively. A comparison of enzyme activities of SDS extracts with those of untreated, control Triton X-100-insoluble membrane residues indicated that both the succinate dehydrogenase and the adenosine triphosphatase antigens were released with a full (or enhanced) catalytic potential at or below concentrations of SDS required to effect maximal solubilization of the enzyme in question. Evidence is also presented to suggest that the more acidic of the two components detected by crossed immunoelectrophoresis for the heterogeneous adenosine triphosphatase antigen is more sensitive to SDS than is the other. Both succinate dehydrogenase and adenosine triphosphatase lost catalytic activity and were denatured at protein/SDS ratios lower than 3.4:1.
D000251:Adenosine Triphosphatases / Q000276:immunology*; D000942:Antigens, Bacterial / Q000032:analysis*; D001426:Bacterial Proteins / Q000276:immunology*; D007123:Immunoelectrophoresis, Two-Dimensional; D007425:Intracellular Membranes / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology*; D008837:Micrococcus / Q000276:immunology*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D013385:Succinate Dehydrogenase / Q000276:immunology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D008565:Membrane Proteins; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D013385:Succinate Dehydrogenase; D000251:Adenosine Triphosphatases