Inhibition of the reconstitution of the haemolytic activity of the first component of human complement by a pepsin-derived fragment of subcomponent C1q.
1. A fragment of subcomponent C1q, which contained all the collagen-like features present in the intact molecule, was isolated by pepsin digestion as described by Reid [Biochem. J. (1976) 155, 5-17]. 2. The pepsin-derived fragment of subcomponent C1q did not bind to antibody-coated erythrocytes under conditions where complete binding of sub-component C1q took place. 3. The peptic fragment blocked the reconstitution of C1 haemolytic activity by competing with intact subcomponent C1q in the utilization of a mixture of the other two subcomponents, C1r and C1s. 4. Reduction and alkylation of the interchain disulphide bonds in the pepsin fragment did not markedly affect its inhibitory effect, whereas heating at 56 degrees C for 30min completely abolished the effect. 5. Lathyritic rat skin collagen and CNBr-derived peptides of pig type II collagen showed no ability to mimic the inhibitory effect of the pepsin fragment when tested over the same concentration range as used for the peptic fragment. 6. The peptic fragment was unable to block efficiently the reconstitution of C1 haemolytic activity unless it was added to the mixture of subcomponents C1r and C1s before the attempt to reconstitute C1 haemolytic activity, in solution, or on the surface of antibody-coated erythrocytes. 7. Evidence was obtained that suggested that subcomponent C1q bound the subcomponent C1r-C1s complex more efficiently when the subcomponent C1q was bound to antibody than when it was free in solution.
D001666:Binding Sites, Antibody; D003094:Collagen / Q000494:pharmacology; D003172:Complement C1 / Q000494:pharmacology; D003174:Complement C1 Inactivator Proteins; D003169:Complement Inactivator Proteins; D004912:Erythrocytes / Q000276:immunology; D006461:Hemolysis; D006358:Hot Temperature; D006801:Humans; D010434:Pepsin A; D010446:Peptide Fragments / Q000494:pharmacology
D003172:Complement C1; D003174:Complement C1 Inactivator Proteins; D003169:Complement Inactivator Proteins; D010446:Peptide Fragments; D003094:Collagen; D010434:Pepsin A