Evasion of Toll-like receptor 2 activation by staphylococcal superantigen-like protein 3

Bardoel, Bart W.; Vos, Ric C. H. de; Bouman, T.; Aerts, Piet C.; Bestebroer, Jovanka; Huizinga, Eric G.; Brondijk, T. Harma C.; Strijp, Jos A. G. van; Haas, Carla J. C. de

doi:10.1007/s00109-012-0926-8

Cited by 86 publications

(86 citation statements)

References 42 publications

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“…The SSL3 and SSL4 genes of S. aureus strain NCTC 8325 (SAOUHSC_00386 and SAOUHSC_00389) were used for construction of truncated proteins SSL3ΔN comprising residues 134-326, SSL4ΔN (residues 79-278), and mutants of SSL3ΔN and SSL4ΔN listed in Table S2. All variants were expressed with a noncleavable N-terminal His 6 -tag in Escherichia coli Rosetta-gami(DE3)pLysS, refolded from insoluble fractions and purified as described (5). Proteins were stored in PBS, and protein purity was determined as >95% by SDS/PAGE.…”

Section: Methodsmentioning

confidence: 99%

“…Ectodomains of mouse (Gln25-Ala588, NM_011905) and human (Lys19-Ala589, NM_003264) TLR2 were transiently expressed with the N-terminal His 6 -StrepII 3 -TEV tag in HEK293-EBNA1-S and HEK293-EBNA-1 cells, respectively (U-Protein Express BV) as described (5). Protein yields were optimized by plasmid titration (35), which indicated that transfections with 10-fold dilutions of expression plasmid in nonexpressing dummy plasmid improved TLR2 production approximately two-to threefold.…”

Section: Methodsmentioning

confidence: 99%

“…Among these secreted virulence factors are the staphylococcal superantigen-like proteins (SSLs), a family of 14 proteins located on two genomic clusters (2)(3)(4). Recently, we and others identified SSL3 as a potent inhibitor of Tolllike receptor 2 (TLR2) (5,6), an innate immunity receptor that is a dominant factor in immune recognition of S. aureus (7)(8)(9)(10).…”

mentioning

confidence: 99%

“…SSL5 inhibits neutrophil extravasation (17,18) and phagocyte function (19,20), SSL7 binds IgA and inhibits complement (21), and SSL10 inhibits IgG1-mediated phagocytosis (22,23), blood coagulation (24), and the chemokine receptor CXCR4 (25). In addition to SSL3, also weak TLR2 inhibitory activity was observed for SSL4 (5), but it remains unknown whether that is its dominant function. This variety of immunomodulatory molecules and functions reflects the importance of the different components of our innate immune system in the defense against S. aureus (26).…”

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confidence: 99%

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Structural basis for inhibition of TLR2 by staphylococcal superantigen-like protein 3 (SSL3)

Koymans

Feitsma

Brondijk

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Toll-like receptors (TLRs) are crucial in innate recognition of invading micro-organisms and their subsequent clearance. Bacteria are not passive bystanders and have evolved complex evasion mechanisms. Staphylococcus aureus secretes a potent TLR2 antagonist, staphylococcal superantigen-like protein 3 (SSL3), which prevents receptor stimulation by pathogen-associated lipopeptides. Here, we present crystal structures of SSL3 and its complex with TLR2. The structure reveals that formation of the specific inhibitory complex is predominantly mediated by hydrophobic contacts between SSL3 and TLR2 and does not involve interaction of TLR2-glycans with the conserved Lewis X binding site of SSL3. In the complex, SSL3 partially covers the entrance to the lipopeptide binding pocket in TLR2, reducing its size by ∼50%. We show that this is sufficient to inhibit binding of agonist Pam 2 CSK 4 effectively, yet allows SSL3 to bind to an already formed TLR2-Pam 2 CSK 4 complex. The binding site of SSL3 overlaps those of TLR2 dimerization partners TLR1 and TLR6 extensively. Combined, our data reveal a robust dual mechanism in which SSL3 interferes with TLR2 activation at two stages: by binding to TLR2, it blocks ligand binding and thus inhibits activation. Second, by interacting with an already formed TLR2-lipopeptide complex, it prevents TLR heterodimerization and downstream signaling.S. aureus | Toll-like receptor | immune evasion | innate immunity | crystal structure

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Structural basis for inhibition of TLR2 by staphylococcal superantigen-like protein 3 (SSL3)

Koymans

Feitsma

Brondijk

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…S. aureus also produces a host of superantigen-like proteins (SSL) that contribute to the immune-evasive properties of these virulent bacteria. In this issue, Bardoel et al [7] show us how staphylococcal SSL3 specifically binds and inhibits TLR-2 activation in neutrophils and monocytes. They show that SSL3 specifically targets TLR-2 and that the binding to TLR-2 is in part sialic acid-dependent.…”

mentioning

confidence: 96%

Staphylococcus aureus, Toll-like receptors, superantigens, and their derivatives

Luft

2012

J Mol Med

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Staphylococcal superantigen‐like proteins interact with human MAP kinase signaling protein ERK2

Dutta

Mukherjee

et al. 2019

FEBS Letters

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This study aimed to identify the intracellular binding partner of a unique class of staphylococcal secreted exotoxins called superantigen‐like proteins (SSL) from human macrophage and keratinocyte cell lysates. Here, we report that SSL1 specifically binds to human extracellular signal‐regulated kinase 2 (hERK2), an important stress‐activated kinase in mitogen‐activated protein kinase signaling pathways. Western blot and in vitro binding studies with recombinant hERK2 confirmed the binding interaction of SSL1, SSL7, and SSL10 with hERK2. Moreover, the SSLs‐hERK2 interaction was validated biochemically by ELISA. Our finding shows that SSLs play a novel role by binding with host cell MAP kinase signaling pathway protein. Understanding the SSL‐hERK2 interaction will also provide a basis for designing SSL‐based peptide inhibitors of hERK2 in cancer therapy.

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Evasion of Toll-like receptor 2 activation by staphylococcal superantigen-like protein 3

Cited by 86 publications

References 42 publications

Structural basis for inhibition of TLR2 by staphylococcal superantigen-like protein 3 (SSL3)

Structural basis for inhibition of TLR2 by staphylococcal superantigen-like protein 3 (SSL3)

Staphylococcus aureus, Toll-like receptors, superantigens, and their derivatives

Staphylococcal superantigen‐like proteins interact with human MAP kinase signaling protein ERK2

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