Localization of the heparin-binding site on complement factor H.
Factor H is a regulator of complement activation and, in this capacity, it prevents activation of the alternative pathway on host cells and tissues when it recognizes markers on these surfaces. This report describes the binding characteristics and location of the site on factor H that is responsible for host recognition. Factor H was found to bind a variety of polyanions, including heparin, heparan sulfate, dextran sulfate, and clusters of sialic acid. In heparin-agarose binding assays it exhibited an affinity for heparin only 2-fold weaker than that of antithrombin III. Factor H exhibited little or no affinity for polyaspartic acid or bacterial colominic acid (polysialic acid). Factor H (Mr 150,000 with approximate dimensions of 30 x 600 A) is composed of 20 highly homologous domains (SCRs) that are arranged as beads on a string. Polyanions were found to block a tryptic cleavage site in domain 15, and a photoaffinity-tagged heparin probe labeled the region between domains 12 and 15. Affinity chromatography of tryptic fragments on heparin-Sepharose confirmed that this region contained the heparin-binding site. CNBr cleavage at Met787 located between SCRs 13 and 14 split the photoaffinity-tagged region. Sequence analysis strongly suggests that domain 13 contains the primary site of polyanion binding. Factor H expresses its complement regulatory function through a site located in domains 4-6 where C3b binds. Thus, the polyanion-binding site that regulates the affinity of factor H for C3b appears to reside more than 200 A away from the C3b-binding site.
Pangburn
M K
MK
Department of Biochemistry, University of Texas Health Science Center, Tyler 75710.
Atkinson
M A
MA
Meri
S
S
D000595:Amino Acid Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D003179:Complement C3b / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000737:chemistry / Q000378:metabolism*; D017242:Complement Factor H; D006493:Heparin / Q000378:metabolism*; D008969:Molecular Sequence Data; D010777:Photochemistry; D012685:Sepharose / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000378:metabolism
D003180:Complement C3b Inactivator Proteins; D014361:Trypsin Inhibitors; D003179:Complement C3b; D017242:Complement Factor H; D006493:Heparin; D012685:Sepharose; D014357:Trypsin