Cell-bound C4b resists reduction by reducing agents: analysis by chain structure and by hemolytic activity.
EA3H-C4hu treated with 2-mercaptoethanol (MSH) or dithiothreitol (DTT) were analyzed for residual chains and hemolytic activity. MSH treatment resulted in no loss of the chains of C4b and no loss of reactivity with a polyclonal anti-human C4 rabbit antibody, and the cells did not lose their ability to generate SAC4b2a. DTT-treated cells lost about 70% of the gamma-chain and over 95% of C4b activity; there was no loss of alpha' and beta chains or the ability to react with anti-C4 antibody. Kinetic studies indicated that the remaining 30% of the gamma-chains could not be removed by prolonged incubation with DTT, implying heterogeneity of cell bound C4b. The data also imply that the gamma-chain is important in the generation of SAC4b2a.
D000818:Animals; D003181:Complement C4 / Q000378:metabolism*; D015935:Complement C4b; D004229:Dithiothreitol / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000378:metabolism; D006168:Guinea Pigs; D006461:Hemolysis; D006801:Humans; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology*; D011487:Protein Conformation; D011817:Rabbits; D051381:Rats; D012756:Sheep
D003181:Complement C4; D008623:Mercaptoethanol; D015935:Complement C4b; D004229:Dithiothreitol