Advertisement

Biotechnology Letters

, Volume 39, Issue 9, pp 1413–1423 | Cite as

Identification of Set11 from Staphylococcus aureus Mu50 strain as a ortholog of SSL7 via bioinformatic analysis and determination of its possible targets from human serum using CNBr-pull down assays

  • Ying Li
  • Xiaobao Chen
  • Nan Jia
  • Xuan ZhangEmail author
  • Jianye Zang
Original Research Paper

Abstract

Objectives

To identify and characterize staphylococcus exotoxin-like (SET) protein Set11 from Staphylococcus aureus Mu50 strain and its possible targets proteins from human blood/serum.

Results

Set11 is a member of the staphylococcal superantigen-like (SSL) proteins (also called Staphylococcus exotoxin-like (SET) proteins) family that is found in staphylococcal strain Mu50. Its structure and function, however, remain unknown. We performed bioinformatics analysis of Set11: it had 90% sequence identity to SSL7 in NCTC 8325 strain, indicating Set11 is a SSL7 ortholog. SSL7 in ATCC 12598 strain binds complement C5 to inhibit complement system. To investigate if Set11 binds C5, we made the homology model of Set11 and the Set11-C5 complex model based on SSL7 and SSL7-C5 structures, respectively. Structural analysis and sequence alignment reveal that the residues in SSL7 involved in C5 binding are conserved in Set11, indicating C5 as the potential target for Set11. To identify new targets of Set11, we cloned, expressed and purified Set11 and performed CNBr-pull down combined mass spectrum assays using human blood and serum.

Conclusions

We identified Set11 as the ortholog of SSL7 and determined C5, fibronectin 1 isoform 3 preproprotein, albumin, alpha-1-microglobulin precursor and complement C3 processor as the potential target proteins for Set11, indicating new functions of Set11/SSL7.

Keywords

Albumin Bioinformatics Exotoxin-like protein C5 Set11 Staphylococcus aureus Structure model SSL7 

Notes

Acknowledgements

This work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No. XDB 08010101), the National Key Research and Development Program of China (Nos. 2016YFA0400903) and the Foundation for Innovative Research Groups of the National Natural Science Foundation of China (Nos. 31621002). This work was also supported by the National Natural Science Foundation of China (Nos. U1532109, 31370756, and 31361163002), and the Scientific Research Grant of Hefei Science Center of CAS (Nos. 2015SRG-HSC043, 2015HSC-UP019) to JZ. This work was also supported by the Anhui Provincial Natural Science Foundation (Nos. 1608085QC52) to XZ and the National Natural Science Foundation of China (Nos. 31400627) to MW.

Supporting information

Supplementary Table 1—Sequences of Set11 and SSL proteins.

Supplementary Table 2—LC-MS/MS results for possible targets of Set11 from human blood/serum.

Supplementary Fig. 1—Structural comparison of Set11 model and SSL7 (PDB ID: 1V1O).

Supplementary Fig. 2—Structure model of Set11-C5.

Supplementary Fig. 3—Signal-peptide prediction of Set11.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10529_2017_2374_MOESM1_ESM.docx (1.7 mb)
Supplementary material 1 (DOCX 1775 kb)

References

  1. Adams PD, Afonine PV, Bunkoczi G, Chen VB et al (2010) PHENIX: a comprehensive python-based system for macromolecular structure solution. Acta Crystallogr D 66:213–221CrossRefPubMedPubMedCentralGoogle Scholar
  2. Bardoel BW, Vos R, Bouman T, Aerts PC, Bestebroer J, Huizinga EG, Brondijk TH, van Strijp JA, de Haas CJ (2012) Evasion of Toll-like receptor 2 activation by staphylococcal superantigen-like protein 3. J Mol Med 90:1109–1120CrossRefPubMedGoogle Scholar
  3. Bestebroer J, Aerts PC, Rooijakkers SHM, Pandey MK, Kohl J, van Strijp JA, de Haas CJ (2010) Functional basis for complement evasion by staphylococcal superantigen-like 7. Cell Microbiol 12:1506–1516CrossRefPubMedPubMedCentralGoogle Scholar
  4. Chung MC, Wines BD, Baker H, Langley RJ, Baker EN, Fraser JD (2007) The crystal structure of staphylococcal superantigen-like protein 11 in complex with sialyl Lewis X reveals the mechanism for cell binding and immune inhibition. Mol Microbiol 66:1342–1355CrossRefPubMedGoogle Scholar
  5. Corpet F (1988) Multiple sequence alignment with hierarchical clustering. Nucleic Acid Res 16:10881–10890CrossRefPubMedPubMedCentralGoogle Scholar
  6. Dutta D, Dutta A, Bhattacharjee A, Basak A, Das AK (2014) Cloning, expression, crystallization and preliminary X-ray diffraction studies of staphylococcal superantigen-like protein 1 (SSL1). Acta Crystallogr F Struct Biol Commun 70:600–603CrossRefPubMedPubMedCentralGoogle Scholar
  7. Fitzgerald JR, Reid SD, Ruotsalainen E, Tripp TJ, Liu M, Cole R, Kuusela P, Schlievert PM, Jarvinen A, Musser JM (2003) Genome diversification in Staphylococcus aureus: molecular evolution of a highly variable chromosomal region encoding the Staphylococcal exotoxin-like family of proteins. Infect Immun 71:2827–2838CrossRefPubMedPubMedCentralGoogle Scholar
  8. Gouet P, Robert X, Courcelle E (2003) ESPript/ENDscript: extracting and rendering sequence and 3D information from atomic structures of proteins. Nucleic Acids Res 31:3320–3323CrossRefPubMedPubMedCentralGoogle Scholar
  9. Itoh S, Hamada E, Kamoshida G, Yokoyama R, Takii T, Onozaki K, Tsuji T (2010) Staphylococcal superantigen-like protein 10 (SSL10) binds to human immunoglobulin G (IgG) and inhibits complement activation via the classical pathway. Mol Immunol 47:932–938CrossRefPubMedGoogle Scholar
  10. Itoh S, Yokoyama R, Kamoshida G, Fujiwara T, Okada H, Takii T, Tsuji T, Fujii S, Hashizume H, Onozaki K (2013) Staphylococcal superantigen-like protein 10 (SSL10) inhibits blood coagulation by binding to prothrombin and factor Xa via their gamma-carboxyglutamic acid (Gla) domain. J Biol Chem 288:21569–21580CrossRefPubMedPubMedCentralGoogle Scholar
  11. Koymans KJ, Vrieling M, Gorham RD Jr, van Strijp JA (2016) Staphylococcal immune evasion proteins: structure, function, and host adaptation. Curr Top Microbiol Immunol. Springer, Berlin Heidelberg, pp 1–49Google Scholar
  12. Kuroda M, Ohta T, Uchiyama I, Baba T et al (2001) Whole genome sequencing of meticillin-resistant Staphylococcus aureus. Lancet 357:1225–1240CrossRefPubMedGoogle Scholar
  13. Langley R, Wines B, Willoughby N, Basu I, Proft T, Fraser JD (2005) The staphylococcal superantigen-like protein 7 binds IgA and complement C5 and inhibits IgA-Fc alpha RI binding and serum killing of bacteria. J Immunol 174:2926–2933CrossRefPubMedGoogle Scholar
  14. Lowy FD (1998) Staphylococcus aureus infections. N Engl J Med 339:520–532CrossRefPubMedGoogle Scholar
  15. McCarthy AJ, Lindsay JA (2013) Staphylococcus aureus innate immune evasion is lineage-specific: a bioinfomatics study. Infect Genet Evol 19:7–14CrossRefPubMedGoogle Scholar
  16. Novick RP (2003) Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol Microbiol 48:1429–1449CrossRefPubMedGoogle Scholar
  17. Patel D, Wines BD, Langley RJ, Fraser JD (2010) Specificity of staphylococcal superantigen-like protein 10 toward the human IgG1 Fc domain. J Immunol 184:6283–6292CrossRefPubMedGoogle Scholar
  18. Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786CrossRefPubMedGoogle Scholar
  19. Schwarz Linek U, Werner JM, Pickford AR, Gurusiddappa S et al (2003) Pathogenic bacteria attach to human fibronectin through a tandem beta-zipper. Nature 423:177–181CrossRefPubMedGoogle Scholar
  20. Williams RJ, Ward JM, Henderson B, Poole S, O’Hara BP, Wilson M, Nair SP (2000) Identification of a novel gene cluster encoding staphylococcal exotoxin-like proteins: characterization of the prototypic gene and its protein product, SET1. Infect Immun 68:4407–4415CrossRefPubMedPubMedCentralGoogle Scholar
  21. Yokoyama R, Itoh S, Kamoshida G, Takii T, Fujii S, Tsuji T, Onozaki K (2012) Staphylococcal superantigen-like protein 3 binds to the Toll-like receptor 2 extracellular domain and inhibits cytokine production induced by Staphylococcus aureus, cell wall component, or lipopeptides in murine macrophages. Infect Immun 80:2816–2825CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Ying Li
    • 1
    • 2
  • Xiaobao Chen
    • 1
    • 2
  • Nan Jia
    • 1
    • 2
  • Xuan Zhang
    • 1
    • 2
    Email author
  • Jianye Zang
    • 1
    • 2
  1. 1.Hefei National Laboratory for Physical Sciences at Microscale CAS Center for Excellence in Biomacromolecules, Collaborative Innovation Center of Chemistry for Life Sciences, and School of Life SciencesUniversity of Science and Technology of ChinaHefeiPeople’s Republic of China
  2. 2.Key Laboratory of Structural BiologyChinese Academy of SciencesHefeiPeople’s Republic of China

Personalised recommendations