Two forms of guinea pig factor B of the alternative complement pathway with different molecular weights.
Two forms of guinea pig factor B (B) of the alternative complement pathway with different mol. wts (Mr) have been isolated from plasma and characterized. The Mr of the two B species, tentatively termed B1 and B2, were estimated to be about 100,000 and 96,000, respectively, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Incubation of B with guinea pig C3 and human factor D (D) in the presence of Mg2+ generated two cleavage fragments of B, namely Ba and Bb. Although the Bb fragments showed the same migration corresponding to an Mr of 62,000, Ba fragments showed different mobilities corresponding to an Mr of 38,000 from B1 and 34,000 from B2. Digestion of B1-Ba, the Ba fragment derived from B1, and B2-Ba, the Ba fragment derived from B2, with endoglycosidase F resulted in a band at Mr 30,000 on an SDS-PAGE in both cases, indicating a difference in structure of the asparagine-linked oligosaccharide moiety in B1-Ba and B2-Ba. No difference in antigenicity was noted between B1 and B2 on immunodiffusion with anti-B sera. Immunoblotting analysis showed that all individual Hartley guinea pigs examined in this study possessed both B1 and B2 at similar levels, as determined by the intensity of staining of their sera. Furthermore, treatment of their serum with zymosan led to the generation of two Ba species corresponding to the Ba fragments from B1 and B2. The capacity to form C3/C5 convertase, as determined by hemolytic assay, was found to be similar between B1 and B2. Furthermore, kinetics of the decay of C3 convertase showed the same half-life of 3.0 min at 30 degrees C. The NH2-terminal amino acid sequences of B1 and B2 and their Bb fragments were determined and found to be identical.
Matsushita
M
M
Department of Microbiology, Fukuoka University School of Medicine, Japan.
Okada
H
H
D000595:Amino Acid Sequence; D000818:Animals; D003167:Complement Activation; D011415:Complement Factor B / Q000145:classification* / Q000276:immunology / Q000302:isolation & purification; D003170:Complement Pathway, Alternative; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000145:classification*; D006168:Guinea Pigs; D006461:Hemolysis; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight
D004792:Enzyme Precursors; D011415:Complement Factor B