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Structural basis for engagement by complement factor H of C3b on a self surface

Nature Structural & Molecular Biology volume 18, pages 463–470 (2011)Cite this article

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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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Figure 1: Introduction to C3b/C3d and complement factor H.
Figure 2: Structure of the C3d–FH19–20 complex showing interacting domains and overall molecular architecture.
Figure 3: Heteronuclear NMR spectroscopy used to map the C3d and dp8 binding sites on FH19–20.
Figure 4: SPR studies of FH19–20 mutants binding to C3d.
Figure 5: Potential model of factor H engagement with surface-bound C3b.
Figure 6: The locations of missense mutations associated with the development of aHUS (and of Q1139A) mapped onto the structure of the C3d–FH19–20 complex.

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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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Author notes

  1. Bärbel S Blaum & Andrew P Herbert

    Present address: Present addresses: Interfaculty Institute for Biochemistry, University of Tübingen, Tübingen, Germany (B.S.B.); The Physiological Laboratory, School of Biomedical Sciences, University of Liverpool, Liverpool, UK (A.P.H.).,

  2. Hugh P Morgan and Christoph Q Schmidt: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Structural and Molecular Biology, School of Biological Sciences, King's Buildings, University of Edinburgh, Edinburgh, UK.,

    Hugh P Morgan, Dominic Gillespie & Jonathan P Hannan

  2. Edinburgh Biomolecular NMR Unit, EaStCHEM, School of Chemistry, University of Edinburgh, Edinburgh, UK

    Christoph Q Schmidt, Mara Guariento, Bärbel S Blaum, Andrew P Herbert, Conny M Johansson, Dušan Uhrín & Paul N Barlow

  3. The Institute of Human Genetics, University of Newcastle upon Tyne, Newcastle upon Tyne, UK

    David Kavanagh

  4. European Molecular Biology Laboratory, Hamburg Outstation, Hamburg, Germany

    Haydyn D T Mertens & Dmitri I Svergun

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Contributions

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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Correspondence to Paul N Barlow or Jonathan P Hannan.

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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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