Identification of the blood coagulation factor interacting sequences in staphylococcal superantigen-like protein 10

Itoh, Saotomo; Takii, Takemasa; Onozaki, Kikuo; Tsuji, Tsutomu; Hida, Shigeaki

doi:10.1016/j.bbrc.2017.02.053

Cited by 10 publications

(16 citation statements)

References 24 publications

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“…Another example is the superantigen-like protein 10 (SSL10) of Staphylococcus aureus, a bacterial exoprotein which directly binds to the Gla-domain of PT and of factor Xa, and binding is competed by Ca 2+ (41). SSL10 has two short and highly basic segments (pI ~11 and pI ~9.8) that are responsible for the binding of SSL10 to PT (42). The interaction likely occurs between the highly positively charged segments and the highly negatively charged Gla-domain of PT.…”

Section: Discussionmentioning

confidence: 99%

Prothrombin is a binding partner of the human receptor of advanced glycation end products

Degani

Altomare

Digiovanni

et al. 2020

Journal of Biological Chemistry

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The receptor for advanced glycation end products (RAGE) plays a key role in mammal physiology and in the etiology and progression of inflammatory and oxidative stress-based diseases. In adults, RAGE expression is normally high only in the lung where the protein concentrates in the basal membrane of alveolar Type I epithelial cells. In diseases, RAGE levels increase in the affected tissues and sustain chronic inflammation. RAGE exists as a membrane glycoprotein with an ectodomain, a transmembrane helix and a short carboxyl-terminal tail, or as a soluble ectodomain that acts as a decoy receptor (sRAGE). VC1 domain is responsible for binding to the majority of RAGE ligands including advanced glycation end products (AGEs), S100 proteins and HMGB1. To ascertain whether other ligands exist, we analyzed by MS the material pulled-down by VC1 from human plasma. Twenty out of 295 identified proteins were selected and associated to coagulation and complement processes and to extracellular matrix. Four of them contained a γ-carboxyl glutamic acid (Gla)-domain, a calcium-binding module, and prothrombin (PT) was the most abundant. Using MicroScale Thermophoresis, we quantified the interaction of PT with VC1 and sRAGE in the absence or presence of calcium that acted as a competitor. PT devoid of the Gla-domain (PT des-Gla) did not bind to sRAGE, providing further evidence that the Gla-domain is critical for the interaction. Finally, the presence of VC1 delayed plasma clotting in a dose-dependent manner. We propose that RAGE is involved in modulating blood coagulation presumably in conditions of lung injury

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Section: Discussionmentioning

confidence: 99%

Prothrombin is a binding partner of the human receptor of advanced glycation end products

Degani

Altomare

Digiovanni

et al. 2020

Journal of Biological Chemistry

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“…3) Chimeras between SSL10 and SSL7, named SSL7x10, SSL10x7 and SSL7x10A-Q, were constructed by domain swapping. 8) SSL7x10R, the mutant of SSL10 replaced β1-β3 to that of SSL7, was constructed by megaprimer method as described previously. 8) The sites where swapped were shown in Fig.…”

Section: Ssl7 Ssl10 and Their Chimerasmentioning

confidence: 99%

“…8) SSL7x10R, the mutant of SSL10 replaced β1-β3 to that of SSL7, was constructed by megaprimer method as described previously. 8) The sites where swapped were shown in Fig. 1, and schematic representations of chimeras were depicted in Fig.…”

Section: Ssl7 Ssl10 and Their Chimerasmentioning

confidence: 99%

“…SSL7, SSL10 and their chimeras were produced in E. coli JM109 as N-terminal hexahistidine-tagged form and purified by Ni-conjugated Sepharose as described previously. 8) The recombinant SSLs were biotinylated using EZ-link NHS-LC-biotin (Thermo Fisher Scientific, Waltham, MA) in according to manufacturer's protocol.…”

Section: Ssl7 Ssl10 and Their Chimerasmentioning

confidence: 99%

“…We previously identified the regions of SSL10 responsible for binding to prothrombin by examining the binding activity of chimeras between SSL10 and its paralog SSL7. 8) We also identified the regions of SSL5 responsible for interaction with its target protein matrix metalloproteinase 9 (MMP-9) by using a series of chimeras between SSL5 and SSL7. 9) In this study, we attempted to identify the regions of SSL10 respon-sible for binding to hIgG by analyzing the binding activity of chimeras to hIgG.…”

Section: Introductionmentioning

confidence: 99%

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Identification of the Regions Responsible for Binding to Human Immunoglobulin G in Staphylococcal Superantigen-Like Protein 10

et al. 2018

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Isolation and functional analysis of phage‐displayed antibody fragments targeting the staphylococcal superantigen‐like proteins

et al. 2023

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Staphylococcus aureus produces numerous virulence factors that manipulate the immune system, helping the bacteria avoid phagocytosis. In this study, we are investigating three immune evasion molecules called the staphylococcal superantigen‐like proteins 1, 5, and 10 (SSL1, SSL5, and SSL10). All three SSLs inhibit vital host immune processes and contribute to S. aureus immune evasion. This study aimed to identify single‐chain variable fragment (scFvs) antibodies from synthetic antibody phage libraries, which can recognize either of the three SSLs and could block the interaction between the SSLs and their respective human targets. The antibodies were isolated after three rounds of panning against SSL1, SSL5, and SSL10, and their ability to bind to the SSLs was studied using a time‐resolved fluorescence‐based immunoassay. We successfully obtained altogether 44 unique clones displaying binding activity to either SSL1, SSL5, or SSL10. The capability of the SSL‐recognizing scFvs to inhibit the SSLs' function was tested in an MMP9 enzymatic activity assay, a P‐selectin glycoprotein ligand 1 competitive binding assay, and an IgG1‐mediated phagocytosis assay. We could show that one scFv was able to inhibit SSL1 and maintain MMP9 activity in a concentration‐dependent manner. Finally, the structure of this inhibiting scFv was modeled and used to create putative scFv‐SSL1‐complex models by protein–protein docking. The complex models were subjected to a 100‐ns molecular dynamics simulation to assess the possible binding mode of the antibody.

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Identification of the blood coagulation factor interacting sequences in staphylococcal superantigen-like protein 10

Cited by 10 publications

References 24 publications

Prothrombin is a binding partner of the human receptor of advanced glycation end products

Prothrombin is a binding partner of the human receptor of advanced glycation end products

Identification of the Regions Responsible for Binding to Human Immunoglobulin G in Staphylococcal Superantigen-Like Protein 10

Isolation and functional analysis of phage‐displayed antibody fragments targeting the staphylococcal superantigen‐like proteins

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