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Characterization and identification of vaccine candidate proteins through analysis of the group A Streptococcus surface proteome

Nature Biotechnology volume 24, pages 191–197 (2006)Cite this article

Abstract

We describe a proteomic approach for identifying bacterial surface-exposed proteins quickly and reliably for their use as vaccine candidates. Whole cells are treated with proteases to selectively digest protruding proteins that are subsequently identified by mass spectrometry analysis of the released peptides. When applied to the sequenced M1_SF370 group A Streptococcus strain, 68 PSORT-predicted surface-associated proteins were identified, including most of the protective antigens described in the literature. The number of surface-exposed proteins varied from strain to strain, most likely as a consequence of different capsule content. The surface-exposed proteins of the highly virulent M23_DSM2071 strain included 17 proteins, 15 in common with M1_SF370. When 14 of the 17 proteins were expressed in E. coli and tested in the mouse for their capacity to confer protection against a lethal dose of M23_DSM2071, one new protective antigen (Spy0416) was identified. This strategy overcomes the difficulties so far encountered in surface protein characterization and has great potential in vaccine discovery.

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Figure 1: The M1_SF370 surface proteome.
Figure 2: Topological analysis of the 37 surface-exposed proteins belonging to the membrane protein family.

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Acknowledgements

We thank J.M. Musser and G. Orefici for providing some of the strains used in this work and Marco Tortoli for his expert assistance in animal studies. We are also grateful to Giorgio Corsi for the artwork and Antonietta Maiorino for her expert secretarial assistance. Manuel J. Rodríguez-Ortega had an Intra-European Marie Curie postdoctoral fellowship from the EU. This work was in part supported by National Institutes of Health/ National Institute of Allergy and Infectious Diseases grant number 5U1060595-02, and in part by the Italian Ministry of Education, University and Research grant no. 10811.

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  1. Chiron Vaccines, Via Fiorentina, Siena, 1 53100, Italy

    Manuel J Rodríguez-Ortega, Nathalie Norais, Giuliano Bensi, Sabrina Liberatori, Sabrina Capo, Marirosa Mora, Maria Scarselli, Francesco Doro, Germano Ferrari, Ignazio Garaguso, Tiziana Maggi, Anita Neumann, Alessia Covre, John L Telford & Guido Grandi

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Correspondence to Guido Grandi.

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All authors work in a private company as either full-time employees or contract scientists and, in particular, G.G. and J.L.T. are holders of company stock options.

Supplementary information

Supplementary Table 1

Surfome of Group A Streptococcus M1_SF370 strain. (PDF 45 kb)

Supplementary Table 2

List of M1_SF370 surfome proteins grouped in degree-of-confidence categories. (PDF 14 kb)

Supplementary Table 3

Surfome of Group A Streptococcus M3_CDCSS90 strain and comparison with the M1_SF370 surfome. (PDF 13 kb)

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Rodríguez-Ortega, M., Norais, N., Bensi, G. et al. Characterization and identification of vaccine candidate proteins through analysis of the group A Streptococcus surface proteome. Nat Biotechnol 24, 191–197 (2006). https://doi.org/10.1038/nbt1179

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