COMPONENT C3 OF HAGFISH COMPLEMENT HAS A UNIQUE STRUCTURE - IDENTIFICATION OF NATIVE C3 AND ITS DEGRADATION PRODUCTS

A protein from hagfish serum that cross-reacted with the third compone nt of hagfish complement (C3) was purified to homogeneity and its stru ctural properties were compared with those of C3 which has a two-subun it chain structure (1 15 and 72 kDa). This protein (designated C3b), w hen purified from plasma, consisted of three disulfide-linked polypept ide chains (77, 72 and 30 kDa). On immunoelectrophoresis, purified C3b migrated more rapidly towards the anode than the beta mobility of C3. However, immunochemical analysis revealed that C3b, after the first s tep in its purification, consisted of two disulfide-linked polypeptide chains (105 and 72 kDa). Treatment of C3b with methylamine, prior to spectrophotometric titration of the free sulfhydryl groups, did not si gnificantly affect the end-point of the titration, suggesting the abse nce of a thioester bond in this molecule. Analysis of the amino acid s equences of the amino-termini of the subunits of C3b revealed that 77 amino acid residues at the amino-terminus of the native cc chain were missing from both the 77-kDa and the 105-kDa polypeptides from C3b. Th ese results indicate that the C3b in this study was analogous to mamma lian C3b. Furthermore, amino acid sequencing data indicated that most of the native C3 from hagfish serum has an irregular two-subunit (alph a + gamma and beta)-linked structure, as a result of one-sided process ing of putative hagfish pro-C3 at the alpha-beta processing site exclu sively. Moreover, it appears that only the molecular features of degen erated hagfish C3 (C3b) are altered during its purification to generat e a three-chain structure.