title: Amidation of C3 at the thiolester site: stimulation of chemiluminescence and phagocytosis by a new inflammatory mediator.
abstract: We studied the ability of particle-bound and fluid-phase C3b monomers and monomeric amidated C3 (prepared by treatment of purified human C3 with ammonium chloride or methylamine) to stimulate chemiluminescence of human polymorphonuclear leukocytes (PMN) and monocytes and to promote phagocytosis in the absence of antibody. Particle-bound C3b evoked chemiluminescence from both PMN and monocytes, and fluid-phase C3b (0.5 mg/ml) elicited significant chemiluminescence from PMN but not from monocytes. Amidated forms of C3, both particle bound and fluid phase, were potent stimulators of chemiluminescence from phagocytic cells and caused a significantly greater response than did C3b. The phagocytosis of 1-micron microspheres by PMN and monocytes was enhanced by coating them with purified C3b in an antibody-free system. Microspheres coated with amidated C3 were avidly phagocytized, and to a greater degree than were C3b-coated microspheres. In a direct binding assay with tritiated monomeric C3b and amidated C3, the affinity of the PMN complement receptor type 1 for C3b (Ka = 4.9 X 10(7) L/M) was similar to that for amidated C3 (Ka = 5.7 X 10(7) L/M). However, there was a fourfold increase in the number of apparent binding sites for amidated C3. This increase did not reflect binding of amidated C3 to the complement receptor type 3, because blocking of CR3 with the monoclonal antibody OKM 10 failed to decrease binding of amidated C3. In sites of increased ammoniagenesis, such as the kidney in chronic renal failure, amidated C3 may play a role as an inflammatory mediator by stimulating oxidative metabolism in phagocytic cells.
mesh_terms: D000643:Ammonium Chloride / Q000494:pharmacology; D001665:Binding Sites; D003176:Complement C3 / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007249:Inflammation / Q000378:metabolism* / Q000503:physiopathology; D007700:Kinetics; D008163:Luminescent Measurements; D008744:Methylamines / Q000494:pharmacology; D008863:Microspheres; D009504:Neutrophils / Q000378:metabolism; D010587:Phagocytosis / Q000187:drug effects
chemical_list: D003176:Complement C3; D008744:Methylamines; D000643:Ammonium Chloride; D003179:Complement C3b; C027451:methylamine