Molecular mechanisms of target recognition in an innate immune system: interactions among factor H, C3b, and target in the alternative pathway of human complement.
In the alternative pathway of complement (APC) factor H is the primary control factor involved in discrimination between potential pathogens. The APC deposits C3b on possible Ags, and the interaction with factor H determines whether the initial C3b activates the APC. Factor H is composed of a linear array of 20 homologous short consensus repeats (SCR) domains with many functional sites. Three of these sites are involved in binding C3b and regulating complement activation; others bind to sialic acid and/or heparin and are responsible for host recognition. Using site-directed mutations we have examined the contributions of each of these sites to target discrimination and to functional activities of factor H. Decay acceleration by SCR1-4 of C3/C5 convertases bound to nonactivators was strongly dependent on SCR domains 11-15 and 16-20. Loss of these regions caused a 97% loss of activity, with SCR16-20 being the most critical (>90% loss). On APC activators the pattern of site usage was different and unique on each. On yeast, deletion of the 10 C-terminal domains (SCR11-20) had no effect on specific activity. On rabbit erythrocytes, this deletion caused loss of 75% of the specific activity. An examination of binding affinity to C3b on the four cell types demonstrated that factor H exhibits a unique pattern of SCR involvement on each cell. The results reveal a complex molecular mechanism of discrimination between microbes and host in this ancient innate defense system and help explain the different rates and intensities of APC activation on different biological particles.
Pangburn
M K
MK
Department of Biochemistry, University of Texas Health Science Center, Tyler, TX 75708, USA. pangburn@uthct.edu
Pangburn
K L
KL
Koistinen
V
V
Meri
S
S
Sharma
A K
AK
D000818:Animals; D018958:CD55 Antigens / Q000378:metabolism; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003179:Complement C3b / Q000378:metabolism*; D017242:Complement Factor H / Q000235:genetics / Q000378:metabolism*; D003170:Complement Pathway, Alternative / Q000235:genetics / Q000276:immunology*; D004910:Erythrocyte Membrane / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D017384:Sequence Deletion; D012756:Sheep
D018958:CD55 Antigens; C493495:CFH protein, human; D011994:Recombinant Proteins; D003179:Complement C3b; D017242:Complement Factor H; D050577:Complement C3-C5 Convertases