1. Proteolytic effects of GelE on complement components in the human serum.
Proteolytic effects of GelE on complement components in the human serum. A, Each reaction mixture was subjected to SDS-PAGE (upper panel) and immunoblotting analysis (lower panel). B, Proteolytic activity of GelE to each purified complement factor. One microgram of each complement factor was treated with GelE at the indicated concentrations for 30 min at room temperature and then subjected to SDS-PAGE. C, The assembly of fluid-phase C3 convertase with fB, fD, and C3b′ generated via GelE action. Validity of C3b′ as a component of C3 convertase was monitored by the appearance of Bb after the incubation of the mixture. Lane 1, human C3b; lane 2, human C3 treated with GelE; lane 3, mixture of C3b, fB, and fD; lane 4, mixture of C3, GelE, fB, and fD. D, Effects of fH and fI on assembled C3 convertase. Lane 1, C3 treated with assembled C3 convertase for control; lane 2, C3 treated with assembled C3 convertase in the presence of fH and fI; lane 3, mixture of C3, GelE, fB, fD, fH, and fI. E, Native SDS-PAGE analysis to examine the fD cofactor activity of C3b and C3b′. Of note, whereas the band for C3 was not changed upon incubation with fB (third lane), those for C3b and C3b′ were moved upward in the presence of fB. In A and E, symbols + and − indicate addition and no addition of corresponding protein into the sample, respectively.
Shin Yong Park et al. J Immunol 2008;181:
Copyright © 2008 by The American Association of Immunologists