Reconstitution of C1 in native proenzyme form and its use in a quantitative C1 activation test.
C1 was reconstituted in macromolecular proenzyme, nonactivated form by incubation of highly purified C1q, C1r, and 125I-C1s together in the presence of calcium. C1 reformed in this manner had an equimolar ratio of C1 subcomponents, as is found in serum, and sedimented in sucrose density gradients with the 16S rate characteristic of C1 in serum. Reconstituted C1 was activatable as shown by cleavage of the 87,000 dalton polypeptide chain of C1s into disulfide linked subunits of 59,000 and 28,000 daltons, respectively, after incubation with aggregated IgG. The extent of activation may be quantitated. Reformed activatable proenzyme C1 can be used to quantitatively assess the C1-activating properties of various substances in addition to its use in the analysis of the C1 activation process.
D002118:Calcium / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003166:Complement Activating Enzymes / Q000378:metabolism*; D003172:Complement C1 / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000378:metabolism*; D006801:Humans; D008970:Molecular Weight
D003172:Complement C1; D004792:Enzyme Precursors; D003166:Complement Activating Enzymes; D002118:Calcium