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Probing the solution structure of Factor H using hydroxyl radical protein footprinting and cross-linking.

TitleProbing the solution structure of Factor H using hydroxyl radical protein footprinting and cross-linking.
Publication TypeJournal Article
Year of Publication2016
AuthorsBaud, A, Gonnet, F, Salard, I, Le Mignon, M, Giuliani, A, Mercère, P, Sclavi, B, Daniel, R
JournalBiochem J
Volume473
Issue12
Pagination1805-19
Date Published2016 06 15
ISSN1470-8728
KeywordsComplement Factor H, Humans, Hydroxyl Radical, Protein Binding, Protein Footprinting, Protein Multimerization, Protein Structure, Tertiary
Abstract

The control protein Factor H (FH) is a crucial regulator of the innate immune complement system, where it is active on host cell membranes and in the fluid phase. Mutations impairing the binding capacity of FH lead to severe autoimmune diseases. Here, we studied the solution structure of full-length FH, in its free state and bound to the C3b complement protein. To do so, we used two powerful techniques, hydroxyl radical protein footprinting (HRPF) and chemical cross-linking coupled with mass spectrometry (MS), to probe the structural rearrangements and to identify protein interfaces. The footprint of C3b on the FH surface matches existing crystal structures of C3b complexed with the N- and C-terminal fragments of FH. In addition, we revealed the position of the central portion of FH in the protein complex. Moreover, cross-linking studies confirmed the involvement of the C-terminus in the dimerization of FH.

DOI10.1042/BCJ20160225
Alternate JournalBiochem. J.
PubMed ID27099340