Chemotactic factor inactivator interaction with Gc-globulin (vitamin D-binding protein). A mechanism of modulating the chemotactic activity of C5a.
Chemotactic factor inactivator (CFI) can decrease the neutrophil chemotactic activity of C5a. Gc-Globulin (GcG) can function as a cochemotaxin for C5a by binding to C5a or C5a des Arg and enhancing its chemotactic potency. We hypothesized that CFI might interact with GcG and thus decrease the chemotactic activity of C5a. CFI was found to markedly inhibit the neutrophil chemotactic activity of partially purified C5a containing GcG (p less than 0.01). Addition of GcG was able to reverse the capacity of CFI to inhibit C5a-directed neutrophil chemotaxis (p less than 0.01). CFI had no significant effect on neutrophil chemotaxis when incubated with C5a depleted of GcG or C5a des Arg. CFI was also able to inhibit the interaction of C5a with GcG adsorbed to plastic. To determine if CFI interacted with GcG, a sandwich ELISA was used. These ELISA tests demonstrated that CFI directly interacted with GcG in a dose-dependent manner that was both heat and pH sensitive. To investigate the possibility of enzymatic degradation of C5a by CFI, CFI preparations were analyzed for carboxypeptidase activity, aminopeptidase activity, and for the capacity to cleave dansylated C5a. No enzymatic activity or cleavage was observed. Furthermore, the direct interaction of CFI with C5a and C5a des Arg was assessed by ELISA tests and column chromatography and no interaction was observed. These results suggest that CFI modulates C5a-directed neutrophil chemotaxis by interacting with GcG and preventing GcG from enhancing the chemotactic potency of C5a.
Robbins
R A
RA
Research Service, Omaha Veterans Administration Medical Center, NE.
Hamel
F G
FG
D000626:Aminopeptidases / Q000378:metabolism; D002268:Carboxypeptidases / Q000378:metabolism; D002630:Chemotactic Factors / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D015936:Complement C5a / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D009504:Neutrophils / Q000502:physiology; D011485:Protein Binding; D014809:Vitamin D-Binding Protein / Q000378:metabolism*
D002630:Chemotactic Factors; D014809:Vitamin D-Binding Protein; C010073:chemotactic factor inactivator; D015936:Complement C5a; D002268:Carboxypeptidases; D000626:Aminopeptidases