Activation of therminal components of human complement by a trypsin-activated complex of human factor B and cobra venom factor.
Cleavage of C3 by CVF-B was demonstrated by hemolytic, immunoelectrophoretic and immune adherence reactions. No cleavage of C5 was detected by immunoelectrophoresis, but C5 hemolytic activity, assayed with EAC1423, decreased although less than C3 hemolytic activity. The co-existence of C3 with limiting amounts of C5 did not reduce the final degree of hemolysis of guinea pig erythrocytes (GPE) induced by late-acting components C6 through C9 and CVF-B. Thus, a CVF-B hemolytic system composed of GPE, C5 through C9 and CVF-B provided a method for titration of terminal components of human complement. CVF-B was able to generate hemolytically active sites of C567 on GPE by activation of C5, C6 and C7. The complex C567 in the fluid-phase decayed within 1 min but C567 on GPE was quite stable. Originally insensitive sheep erythrocytes became sensitive to the CVF-B hemolytic system if C3b sites were present, suggesting that cell-bound C3b played a role in orienting the positions of C567 to be fixed. CVF-B could be recovered quantitatively from the supernatant of the reaction mixture in which the hemolytically active intermediate GPEC-5678 had been formed through the interaction between C5 to C8 and CVF-B.
D000818:Animals; D003176:Complement C3 / Q000378:metabolism; D003182:Complement C5 / Q000378:metabolism; D003183:Complement C6 / Q000378:metabolism; D003184:Complement C7 / Q000378:metabolism; D003186:Complement C9 / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism*; D006168:Guinea Pigs; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D007106:Immune Sera; D066298:In Vitro Techniques; D012910:Snake Venoms / Q000494:pharmacology*; D014357:Trypsin / Q000494:pharmacology*
D003176:Complement C3; D003182:Complement C5; D003183:Complement C6; D003184:Complement C7; D003186:Complement C9; D007106:Immune Sera; D012910:Snake Venoms; D003165:Complement System Proteins; D014357:Trypsin