Detection of two variants of complement component C3 in C3-deficient guinea pigs distinguished by the absence and presence of a thiolester.
The complement system is an essential part of the innate defense, and C3 is an integral part of this powerful system. In previously identified complement C3 deficient guinea pigs only approx. 5% of the normal serum C3 level is detectable. No differences were found between in vitro C3 protein synthesis and C3 mRNA levels of cells from C3-deficient and wild-type animals and the amino acid sequences of both C3 proteins are identical as deduced from cDNA sequencing. Previously, the principal inability to form a C3 thiolester was discussed as a possible reason for this C3-deficiency. Here we report the isolation of two functionally different C3 species from the C3-deficient animals. Only one of these C3 proteins exhibits normal hemolytic activity and contains a thiolester group. The second C3 species is exclusively present in C3-deficient animals and lacks a thiolester, explaining its failure to express hemolytic activity. The presence of a second C3 species lacking a thiolester structure only in C3-deficient animals indicates that the stability of the thiolester may play a role in C3 deficiency. However further analysis of the in vitro stability of the thiolesters of C3 from normal and C3-deficient guinea pigs revealed no differences. A decreased in vivo thiolester stability might lead to the presence of C3 with and without a thiolester or alternatively the expression of two isoforms of C3 in these animals. Considering the central role of C3 in host defense, the mechanisms of C3 thiolester formation require further analysis.
Lamping
N
N
Robert-Koch Institut, Nordufer 20, 13353, Berlin, Germany.
Schumann
R R
RR
Burger
R
R
D000818:Animals; D001710:Biotin; D015153:Blotting, Western / Q000379:methods; D002241:Carbohydrates / Q000032:analysis; D002250:Carbon Radioisotopes; D005591:Chemical Fractionation; D003176:Complement C3 / Q000032:analysis* / Q000145:classification / Q000276:immunology; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006168:Guinea Pigs; D006461:Hemolysis; D007553:Isotope Labeling; D008744:Methylamines; D051379:Mice; D018345:Mice, Knockout; D020033:Protein Isoforms / Q000032:analysis / Q000145:classification / Q000276:immunology; D012967:Sodium Dodecyl Sulfate; D013438:Sulfhydryl Compounds / Q000032:analysis*
D002241:Carbohydrates; D002250:Carbon Radioisotopes; D003176:Complement C3; D008744:Methylamines; D020033:Protein Isoforms; D013438:Sulfhydryl Compounds; D003208:Concanavalin A; D012967:Sodium Dodecyl Sulfate; D001710:Biotin