4.4 Article

Interaction between the Staphylococcus aureus extracellular adherence protein Eap and its subdomains with platelets

Journal

INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY
Volume 308, Issue 6, Pages 683-691

Publisher

ELSEVIER GMBH
DOI: 10.1016/j.ijmm.2018.04.004

Keywords

Staphylococcus aureus; Platelets; Extracellular adherence protein; Micropattern arrays; Bacteriomimetic microbeads

Funding

  1. German Research Foundation [DFG CRC TRR34]

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S. aureus associated bacteremia can lead to severe infections with high risk of mortality (e.g. sepsis, infective endocarditis). Many virulence factors and adhesins of S. aureus are known to directly interact with platelets. Extracellular adherence protein, Eap, one of the most important virulence factors in S. aureus mediated infections is a multi-tandem domain protein and has been shown to interact with almost all cell types in the human circulatory system. By using amine reactive fluorescent N-hydroxysuccinimidyl (NHS)-ester dyes and by direct detection with primary fluorescently conjugated anti-histidine (His-tag) antibodies against detect N-terminal His(6), we show Eap subdomain Eap D3D4 specifically interacts and rapidly activates human platelets. Furthermore, we validate our finding by using site directed directional immobilization of Eap D3D4 through N-terminal His(6) on nickel (II)-nitrilotriacetic acid (Ni-NTA) functionalized bacteriomimetic microbead arrays to visualize real-time platelet activation through calcium release assay. These methods offer an easily adoptable protocols for screening of S.aureus derived virulence factors and adhesins with platelets.

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