title: The immunochemical approach to the characterization of membrane proteins. Human erythrocyte membrane proteins analysed as a model system.
abstract: 1. Crossed immunoelectrophoresis was used for extensive characterization of individual proteins of human erythrocyte membranes solubilized in non-ionic detergent. 2. The precipitates were assigned to extrinsic or intrinsic proteins. 3. Four glycoproteins were identified by their lectin binding behaviour, whilst five proteins were affected by neuraminidase, indicating them to be sialoglycoproteins. 4. Enzymatic activity is retained in the solubilized system and the presence of acetylcholinesterase and an ATPase was demonstrated. The formation of phosphorylated membrane proteins on incubation with [32P]ATP was demonstrated by autoradiography on the immunoelectrophoresis plates. 5. Five proteins located on the outer cell surface were identified by antibody binding to intact cells. These same proteins were degraded by proteolytic enzymes in intact cells but only three of them were labelled by lactoperoxidase-catalysed 125I-iodination. 6. Analysis of erythrocyte membrane proteins using quantitive immunoelectrophoresis yields results concordant with those obtained by dodecyl sulfate-polyacrylamide gel electrophoresis.
mesh_terms: D001665:Binding Sites; D004910:Erythrocyte Membrane / Q000648:ultrastructure; D004912:Erythrocytes / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods; D007784:Lactoperoxidase; D037102:Lectins; D008565:Membrane Proteins / Q000097:blood*; D008954:Models, Biological; D009994:Osmolar Concentration; D011485:Protein Binding; D012795:Sialoglycoproteins / Q000097:blood
chemical_list: D037102:Lectins; D008565:Membrane Proteins; D012795:Sialoglycoproteins; D007784:Lactoperoxidase