The structural basis for inhibition of the classical and lectin complement pathways by S. aureus extracellular adherence protein

Protein Science : a Publication of the Protein Society
Jordan L WoehlBrian V Geisbrecht

Abstract

The extracellular adherence protein (Eap) plays a crucial role in pathogenesis and survival of Staphylococcus aureus by inhibiting the classical and lectin pathways of complement. We have previously shown that Eap binds with nanomolar affinity to complement C4b and disrupts the initial interaction between C4b and C2, thereby inhibiting formation of the classical and lectin pathway C3 pro-convertase. Although an underlying mechanism has been identified, the structural basis for Eap binding to C4b is poorly understood. Here, we show that Eap domains 3 and 4 each contain a low-affinity, but saturable binding site for C4b. Taking advantage of the high lysine content of Eap, we used a zero-length crosslinking approach to map the Eap binding site to both the α'- and γ-chains of C4b. We also probed the C4b/Eap interface through a chemical footprinting approach involving lysine modification, proteolytic digestion, and mass spectrometry. This identified seven lysines in Eap that undergo changes in solvent exposure upon C4b binding. We found that simultaneous mutation of these lysines to either alanine or glutamate diminished C4b binding and complement inhibition by Eap. Together, our results provide insight into Eap recognition of C4b, ...Continue Reading

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Citations

Dec 20, 2018·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Stefania GiussaniImmaculada Margarit
Feb 8, 2019·Biomolecular NMR Assignments·Alvaro I HerreraOm Prakash
Nov 9, 2017·Protein Science : a Publication of the Protein Society·Daphne A C StapelsBrian V Geisbrecht

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