Analysis of recognition in the alternative pathway of complement. Effect of polysaccharide size.
Covalent attachment of the complement (C) protein C3b to polysaccharides on biologic particles which activate the alternative pathway leads to changes in the affinity of C3b for factor H, a regulatory protein of the C system. In this study the size of the site with which the polysaccharides interact and its special relationship to the thioester site were investigated using a fluorimetric assay and soluble C3b attached to low m.w. polysaccharides. Oligomers of alpha 1-6 and alpha 1-4 polyglucose and beta 1-2 polyfructose were prepared and attached to C3b at the thioester site. C3b bound to monomeric, dimeric, or trimeric sugars exhibited the same interaction with factor H as free C3b, i.e., there was no effect due to attachment alone. Beginning with tetrameric oligosaccharides a linear decrease in factor H binding was observed with increasing oligosaccharide size and the effect reached an apparent maximum with large polysaccharides. Maximum inhibition of factor H function was estimated to occur at a length of 16 saccharide units. The results suggest that this site, which regulates the inactivation rate of surface-bound C3b and thus the activation of the alternative pathway of C, spans a maximum of 13 sugar units (less than 65 A) starting four units (approximately 15 A) from the thioester site in C3b.
Pangburn
M K
MK
Department of Biochemistry, University of Texas Health Center, Tyler 75710.
D001665:Binding Sites; D003167:Complement Activation; D003179:Complement C3b / Q000378:metabolism*; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D017242:Complement Factor H; D003170:Complement Pathway, Alternative; D003911:Dextrans / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis; D011134:Polysaccharides / Q000378:metabolism*; D013329:Structure-Activity Relationship
C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D003911:Dextrans; D008070:Lipopolysaccharides; D009844:Oligosaccharides; D011134:Polysaccharides; D003179:Complement C3b; D017242:Complement Factor H