Covalent binding and hemolytic activity of complement proteins.
We report the inactivation of the third component of complement (C3) by hydroxylamine. C3 hemolytic and covalent binding activities decline with identical kinetics, demonstrating a direct correlation between the two activities. We conclude that covalent, surface-bound C3b is hemolytically active. The inactivation of C3 is first order with respect to hydroxylamine. We also studied C3 inactivation with [14C]methylamine. The inactivation corresponds quantitatively with the labeling of C3 in the C3d domain. The data obtained support the following hypothesis: there is an internal thioester within C3 which becomes highly reactive on activation to C3b, and C3b binds to receptive surfaces by transfer of the acyl function of the thioester to a hydroxyl group on the receptive surface. This proposed model for the reaction of C3 with receptive surfaces also applies to C4, which binds to membrane surfaces covalently and is able to be inactivated by hydroxylamine and methylamine. C5, on the other hand, is not inactivated by treatment with the amines.
D001665:Binding Sites / Q000187:drug effects; D003176:Complement C3 / Q000037:antagonists & inhibitors*; D003181:Complement C4 / Q000037:antagonists & inhibitors; D003182:Complement C5 / Q000037:antagonists & inhibitors; D004952:Esters; D006461:Hemolysis; D006898:Hydroxylamines / Q000494:pharmacology*; D007700:Kinetics; D008744:Methylamines / Q000494:pharmacology
D003176:Complement C3; D003181:Complement C4; D003182:Complement C5; D004952:Esters; D006898:Hydroxylamines; D008744:Methylamines