Abstract
Complement factor H (FH) attenuates C3b molecules tethered by their thioester domains to self surfaces and thereby protects host tissues. Factor H is a cofactor for initial C3b proteolysis that ultimately yields a surface-attached fragment (C3d) corresponding to the thioester domain. We used NMR and X-ray crystallography to study the C3d–FH19–20 complex in atomic detail and identify glycosaminoglycan-binding residues in factor H module 20 of the C3d–FH19–20 complex. Mutagenesis justified the merging of the C3d–FH19–20 structure with an existing C3b–FH1–4 crystal structure. We concatenated the merged structure with the available FH6–8 crystal structure and new SAXS-derived FH1–4, FH8–15 and FH15–19 envelopes. The combined data are consistent with a bent-back factor H molecule that binds through its termini to two sites on one C3b molecule and simultaneously to adjacent polyanionic host-surface markers.
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Acknowledgements
Use of the Protein Production Facilities at the University of Edinburgh was supported by The Wellcome Trust, the Scottish University Life Sciences Alliance and the UK Biotechnology and Biological Sciences Research Council. C.Q.S. and M.G. were supported by Wellcome Trust grant 081179, A.P.H. was supported by Wellcome Trust grant 078780/Z/05/Z (to D.U. and P.N.B.), B.S.B. was supported by an EastChem PhD studentship and a University of Edinburgh campaign small project grant and C.M.J. was supported by EC FP6 (Marie Curie EST Fellowship, contract MEST-CT-2005-020744).
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M.G., C.Q.S., D.K. and A.P.H. engineered and produced factor H proteins; J.P.H. and D.G. engineered and produced C3d proteins; C.M.J. generated dp8 heparin fragments. J.P.H. and D.G. crystallized the C3d–FH complex; H.P.M. and J.P.H. collected crystallographic data; H.P.M. determined and refined the structure; B.S.B. produced 15N-labeled factor H and carried out NMR studies of the FH–C3d and FH–dp8 interactions; B.S.B. and A.P.H. carried out the hemolysis assay; J.P.H. carried out the C3d-dp8 competition ELISA; C.Q.S. and M.G. carried out the SPR measurements; H.D.T.M. and D.I.S. carried out the SAXS analysis; D.U., J.P.H. and P.N.B. conceived and supervised the project; H.P.M., C.Q.S., M.G., B.S.B., D.U., J.P.H. and P.N.B. wrote the manuscript.
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Morgan, H., Schmidt, C., Guariento, M. et al. Structural basis for engagement by complement factor H of C3b on a self surface. Nat Struct Mol Biol 18, 463–470 (2011). https://doi.org/10.1038/nsmb.2018
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DOI: https://doi.org/10.1038/nsmb.2018
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