Cyanate as an inactivator of complement proteins.
Sodium cyanate added to normal human serum or serum from patients with sickle-cell disease resulted in the functional inactivation of C3, C5, C6, C7, and the C3b inactivator, but not C8 and C9. Final concentrations as low as 0.5 mM in serum caused inactivation of 12 to 64% of the C3 after 8 hr at 37 degrees C. The activity of the inactivated C3, C5, and C3b inactivator was not restored by dialysis. Most of the functional activity of C3 in cyanate-treated sera was destroyed by very small quantities of 14C-labeled cyanate that was bound to the protein. C3 inactivation by cyanate occurred in heated sera (50 degrees C, 30 min) and sera treated with EDTA, probably indicating that one mechanism for inactivation was by a direct carbamylation reaction. Both C3 and C5 showed two anodal-migrating forms in two dimensional antigen-antibody crossed electrophoresis in some sera treated with low concentrations of cyanate. Measurements of circular dichroism of highly purified carbamylated C3 showed no detectable changes in structure even though most of the functional activity was destroyed. Purified, inactive C3 that was carbamylated with 14C-labeled cyanate was capable of binding to EAC142, but the resulting EAC1423 was weakly positive for immune adherence and negative for agglutination with anti-C3 antiserum. Unlabeled, cell-bound C3b on EAC142 was not susceptible to cyanate action as shown by no loss in immune adherence and positive agglutination with anti-C3 antiserum. The C3b inactivator was more susceptible to cyanate than C3 in a short time period, whereas both were inactivated after 8 hr. Since cyanate is currently being evaluated as a treatment for sickle-cell disease, the inactivation of C3 by the drug is an important consideration for such patients who are already deficient in C3 dependent heat-labile opsonins that aid in host defense.
D000755:Anemia, Sickle Cell / Q000276:immunology; D003176:Complement C3 / Q000037:antagonists & inhibitors; D003182:Complement C5 / Q000037:antagonists & inhibitors; D003183:Complement C6 / Q000037:antagonists & inhibitors; D003184:Complement C7 / Q000037:antagonists & inhibitors; D003185:Complement C8 / Q000037:antagonists & inhibitors; D003186:Complement C9 / Q000037:antagonists & inhibitors; D003169:Complement Inactivator Proteins; D003165:Complement System Proteins / Q000378:metabolism; D003485:Cyanates / Q000494:pharmacology*; D006461:Hemolysis; D006801:Humans; D007104:Immune Adherence Reaction; D007123:Immunoelectrophoresis, Two-Dimensional; D011487:Protein Conformation / Q000187:drug effects
D003176:Complement C3; D003182:Complement C5; D003183:Complement C6; D003184:Complement C7; D003185:Complement C8; D003186:Complement C9; D003169:Complement Inactivator Proteins; D003485:Cyanates; D003165:Complement System Proteins