Electrophoretic characterization of muscarinic receptors under denaturating and nondenaturating conditions: computer-assisted Ferguson plot analysis.
The physical properties of the covalently labeled [( 3H]propylbenzilycholine mustard) muscarinic acetylcholine receptor from rat brain were studied by sodium dodecyl sulfate-polyacrylamide electrophoresis and computer-assisted Ferguson plot analysis. No proteolytic degradation or dimerization of the ligand binding subunit was found. No clues for different molecular weight forms or anomalous migration of the muscarinic receptor were detected. The weighted regression analysis of Ferguson plots gave an apparent molecular mass of 64-65 kDa. A new method for the electrophoretic separation of native (quinuclidinyl[3H]benzilate labeled) muscarinic receptor-detergent complexes was used for the comparison of wheat germ agglutinin binding, and not lectin binding receptors which were obtained by selective solubilization from porcine striatum. For this purpose, the computer-assisted Ferguson plot analysis is particularly suitable, since it renders possible the statistical assessment of both size and charge differences. Both receptor-detergent complexes were found to differ; statistically significant in their net charge but not in their size. The data support the view that muscarinic receptors from different sources may differ considerably in their glycosylation and that the receptor from porcine striatum can reversibly associate with a low-molecular-mass component which contains sialic acid.
D000818:Animals; D001923:Brain Chemistry; D003201:Computers; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008970:Molecular Weight; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains; D011975:Receptors, Mitogen / Q000302:isolation & purification; D011976:Receptors, Muscarinic / Q000302:isolation & purification* / Q000378:metabolism; D012044:Regression Analysis; D013552:Swine; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D003902:Detergents; D011975:Receptors, Mitogen; D011976:Receptors, Muscarinic; D014909:Wheat Germ Agglutinins; C027503:wheat germ agglutinin receptor