Purification from guinea pig erythrocyte stroma of a decay-accelerating factor for the classical c3 convertase, C4b,2a.
A protein with decay-accelerating activity for the classical C3 convertase, C4b2a, has been isolated on the basis of this function from guinea pig erythrocyte stroma. The isolation procedure for decay-accelerating factor of stroma (DAF-S) utilizes butanol extraction and chromatography on DEAE-Sephacel, hydroxylapatite, and phenyl Sepharose. Purified DAF-S has a m.w. of 60,000 and 65,000 on reduced and unreduced SDS gels, respectively, and exhibits m.w. of 30,000 and 175,000 on alkaline gradient gels, suggesting multiples of a 60,000-65,000 subunit. Purified DAF-S elicited a monospecific antiserum whose IgG fraction neutralized the decay-accelerating activity for C4b,2a affixed to 10(7) sheep erythrocytes (EAC1,4,2) in a dose-response fashion. The monospecific antiserum diluted up to 1:5120 agglutinated 1 x 10(6) guinea pig erythrocytes, but not sheep or human erythrocytes, suggesting that DAF-S, an integral membrane protein, has species-specific antigens that are expressed on the surface of the guinea pig erythrocyte.
D000818:Animals; D003166:Complement Activating Enzymes; D003175:Complement C2; D050577:Complement C3-C5 Convertases; D003181:Complement C4; D004912:Erythrocytes / Q000276:immunology*; D006168:Guinea Pigs; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D011817:Rabbits; D013997:Time Factors
D003175:Complement C2; D003181:Complement C4; D007106:Immune Sera; D003166:Complement Activating Enzymes; D050577:Complement C3-C5 Convertases