Generation of the bioactive kallikrein-derived fragment, C3d-k, by HANE-plasma

Recent studies have concluded that after complement activation the final physiologic degradation products of C3 and C3c and the fragment of relative molecular mass (M(r)) 42,000 which contains the C3d and C3g domains and was therefore named C3d,g. Using fluorescent labelled C3b as a substrate, we have determined the putative C3d,g ('C3d,g') producing activity of both normal and hereditary angioneurotic oedema (HANE) plasma. In normal plasmas, the rate of production of C3d,g was 1.0 ± 0.2 x 10^-10 mol/ml/h and this activity was blocked by antibodies to I. In contrast, HANE, plasmas (deficient in C1INH) showed more than twice as much 'C3d,g' production as normal plasmas and both antibodies to I and kallikrein were required to inhibit this activity. Because of this result, a more sensitive gel system was employed to detect the M(r) 42,000 peptide and two 'C3d,g' fragments of approximately equal intensity with M(r) of 42,000 and 43,000 were defined. Incubation of purified kallikrein with labelled iC3b produced a C3d,g-like fragment, C3d-k, that aligned with the band of 43,000 M(r) generated in HANE plasma. These results indicate that HANE plasma, in contrast to normal plasma, generates the bioactive C3d-k fragment. C1INH blocks the activities of kallikrein and C1s, and C3d-k generation in HANE plasma is probably secondary to the proteolytic activity of kallikrein.