title: Disease-associated N-terminal complement factor H mutations perturb cofactor and decay-accelerating activities.
abstract: Many mutations associated with atypical hemolytic uremic syndrome (aHUS) lie within complement control protein modules 19-20 at the C terminus of the complement regulator factor H (FH). This region mediates preferential action of FH on self, as opposed to foreign, membranes and surfaces. Hence, speculation on disease mechanisms has focused on deficiencies in regulation of complement activation on glomerular capillary beds. Here, we investigate the consequences of aHUS-linked mutations (R53H and R78G) within the FH N-terminal complement control protein module that also carries the I62V variation linked to dense-deposit disease and age-related macular degeneration. This module contributes to a four-module C3b-binding site (FH1-4) needed for complement regulation and sufficient for fluid-phase regulatory activity. Recombinant FH1-4(V62) and FH1-4(I62) bind immobilized C3b with similar affinities (K(D) = 10-14 μM), whereas FH1-4(I62) is slightly more effective than FH1-4(V62) as cofactor for factor I-mediated cleavage of C3b. The mutant (R53H)FH1-4(V62) binds to C3b with comparable affinity (K(D) ∼12 μM) yet has decreased cofactor activities both in fluid phase and on surface-bound C3b, and exhibits only weak decay-accelerating activity for C3 convertase (C3bBb). The other mutant, (R78G)FH1-4(V62), binds poorly to immobilized C3b (K(D) >35 μM) and is severely functionally compromised, having decreased cofactor and decay-accelerating activities. Our data support causal links between these mutations and disease; they demonstrate that mutations affecting the N-terminal activities of FH, not just those in the C terminus, can predispose to aHUS. These observations reinforce the notion that deficiency in any one of several FH functional properties can contribute to the pathogenesis of this disease.
authors.affiliation: School of Chemistry, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
authors.forename: Isabell C
authors.initials: IC
authors.lastname: Pechtl
authors.forename: David
authors.initials: D
authors.lastname: Kavanagh
authors.forename: Nicola
authors.initials: N
authors.lastname: McIntosh
authors.forename: Claire L
authors.initials: CL
authors.lastname: Harris
authors.forename: Paul N
authors.initials: PN
authors.lastname: Barlow
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D065766:Atypical Hemolytic Uremic Syndrome; D003179:Complement C3b / Q000235:genetics / Q000378:metabolism; D017242:Complement Factor H / Q000235:genetics / Q000378:metabolism*; D030342:Genetic Diseases, Inborn / Q000235:genetics / Q000378:metabolism*; D006463:Hemolytic-Uremic Syndrome; D006801:Humans; D020125:Mutation, Missense; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012756:Sheep
chemical_list: D011994:Recombinant Proteins; D003179:Complement C3b; D017242:Complement Factor H