Comparison of surface recognition and C3b binding properties of mouse and human complement factor H.
Factor H (FH) is a central complement regulator both in plasma and on certain cellular and acellular surfaces that are in contact with plasma. Although FH deficiency has been shown to lead to similar diseases in man and mice (membranoproliferative glomerulonephritis or dense deposit disease) little is known about the similarity between the human and murine FH functions. We here characterize the interactions of murine FH (mFH) with C3b, glycosaminoglycans, and endothelial cells and compare these interactions with those of human FH (hFH). To achieve this we purified mFH and murine C3 from plasma, prepared murine C3b, and expressed recombinant mFH constructs containing domains 1-5 and 18-20 (mFH1-5 and mFH18-20). For comparisons, hFH, human C3b, and recombinant hFH1-5 and hFH18-20 were used. We demonstrate that mFH and mFH1-5 do act as cofactors for factor I-mediated cleavage of human C3b. Surface plasmon resonance analysis showed binding of mFH18-20 to murine C3b and weak binding to human C3b. The mFH18-20 construct bound to heparin in a manner comparable to hFH18-20. It was demonstrated by flow cytometry that mFH and mFH18-20 bind to human endothelial cells in a similar manner to hFH and hFH18-20. Taken together, locations of the key functions of mFH, i.e. complement regulation and surface recognition, are comparable to hFH. Recently, mutations in the carboxy-terminal end of hFH have been found to be associated with atypical hemolytic uremic syndrome (aHUS). Based on the results in this report it is conceptually attractive to establish a murine model for aHUS.
Cheng
Zhu-Zhu
ZZ
Department of Bacteriology and Immunology, Haartman Institute and HUSLAB, University and University Hospital of Helsinki, Haartmaninkatu 3, FIN-00290 Helsinki, Finland.
Hellwage
Jens
J
Seeberger
Harald
H
Zipfel
Peter F
PF
Meri
Seppo
S
Jokiranta
T Sakari
TS
D000818:Animals; D003179:Complement C3b / Q000737:chemistry / Q000378:metabolism*; D017242:Complement Factor H / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000276:immunology; D006493:Heparin / Q000737:chemistry; D006801:Humans; D051379:Mice; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance
D011994:Recombinant Proteins; D003179:Complement C3b; D017242:Complement Factor H; D006493:Heparin