Mutations of factor H impair regulation of surface-bound C3b by three mechanisms in atypical hemolytic uremic syndrome.
Atypical hemolytic uremic syndrome (aHUS) is a thrombotic microangiopathy associated with mutations in complement proteins, most frequently in the main plasma alternative pathway regulator factor H (FH). The hotspot for the FH mutations is in domains 19-20 (FH19-20) that are indispensable for FH activity on C3b bound covalently to host cells. In aHUS, down-regulation of cell-bound C3b by FH is impaired, but it is not clear whether this is due to an altered FH binding to surface-bound C3b or to cell surface structures. To explore the molecular pathogenesis of aHUS we tested binding of 14 FH19-20 point mutants to C3b and its C3d fragment, mouse glomerular endothelial cells (mGEnC-1), and heparin. The cell binding correlated well, but not fully, with heparin binding and the cell binding site was overlapping but distinct from the C3b/C3d binding site that was shown to extend to domain 19. Our results show that aHUS-associated FH19-20 mutants have different combinations of three primary defects: impaired binding to C3b/C3d, impaired binding to the mGEnC-1 cells/heparin, and, as a novel observation, an enhanced mGEnC-1 cell or heparin binding. We propose a model of the molecular pathogenesis of aHUS where all three mechanisms lead eventually to impaired control of C3b on the endothelial cell surfaces. Based on the results with the aHUS patient mutants and the overlap in FH19-20 binding sites for mGEnC-1/heparin and C3b/C3d we conclude that binding of FH19-20 to C3b/C3d is essential for target discrimination by the alternative pathway.
Lehtinen
Markus J
MJ
Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, FIN-00290 Helsinki, Finland. markus.j.lehtinen@helsinki.fi
Rops
Angelique L
AL
Isenman
David E
DE
van der Vlag
Johan
J
Jokiranta
T Sakari
TS
D019943:Amino Acid Substitution; D000818:Animals; D003179:Complement C3b / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015933:Complement C3d / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017242:Complement Factor H / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017931:DNA Primers; D042783:Endothelial Cells / Q000502:physiology; D006463:Hemolytic-Uremic Syndrome / Q000235:genetics*; D006801:Humans; D007249:Inflammation / Q000503:physiopathology; D007678:Kidney Glomerulus / Q000502:physiology; D007700:Kinetics; D051379:Mice; D016296:Mutagenesis; D009154:Mutation; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D017931:DNA Primers; D011994:Recombinant Proteins; D003179:Complement C3b; D015933:Complement C3d; D017242:Complement Factor H