Abstract
This chapter describes a protocol for the separation of different cell fractions from Streptococcus pyogenes and sample preparation for analysis by mass spectrometry. The presented approach can be used for the analysis of all subcellular proteomes from S. pyogenes and enables identification of drug or vaccine targets within a certain cellular fraction. Here, proteins integral to the plasma membrane are of particular interest for the development of new antimicrobial therapies.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Navarre WW, Schneewind O (1999) Surface proteins of gram-positive bacteria and mechanisms of their targeting to the cell wall envelope. Microbiol Mol Biol Rev 63(1):174–229
Silhavy TJ, Kahne D, Walker S (2010) The bacterial cell envelope. Cold Spring Harb Perspect Biol 2(5):a000414
Scott JR, Barnett TC (2006) Surface proteins of gram-positive bacteria and how they get there. Annu Rev Microbiol 60:397–423
Fischetti VA (2016) M protein and other surface proteins on streptococci. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK. https://www.ncbi.nlm.nih.gov/books/NBK333431/. Accessed 1 June 2017
Xie K, Dalbey RE (2008) Inserting proteins into the bacterial cytoplasmic membrane using the sec and YidC translocases. Nat Rev Microbiol 6(3):234–244
Callegari EA, Chaussee MS (2016) The streptococcal proteome. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK. https://www.ncbi.nlm.nih.gov/books/NBK333416/. Accessed 1 June 2017
Rodriguez-Ortega MJ, Norais N, Bensi G et al (2006) Characterization and identification of vaccine candidate proteins through analysis of the group A Streptococcus surface proteome. Nat Biotechnol 24(2):191–197
Malmström J, Karlsson C, Nordenfelt P et al (2012) Streptococcus pyogenes in human plasma adaptive mechanisms analyzed by mass spectrometry-based proteomics. J Biol Chem 287(2):1415–1425
Sharma A, Arya DK, Sagar V et al (2013) Identification of potential universal vaccine candidates against group a streptococcus by using high throughput in silico and proteomics approach. J Proteome Res 12(1):336–346
Lei B, Mackie S, Lukomski S, Musser JM (2000) Identification and immunogenicity of group A Streptococcus culture supernatant proteins. Infect Immun 68(12):6807–6818
Cole JN, Djordjevic SP, Walker MJ (2008) Isolation and solubilization of Gram-positive bacterial cell wall-associated proteins. In: Posch A (ed) 2D PAGE: sample preparation and fractionation. Methods in molecular biology, vol 425. Humana Press, New York, NY, pp 295–311
Wilk L, Happonen L, Malmström J, Herwald H (2018) Comprehensive mass spectrometric survey of Streptococcus pyogenes subcellular proteomes. J Proteome Res 17(1):600–617
Karlsson C, Teleman J, Malmström J (2017) Analysis of bacterial surface interactions with mass spectrometry-based proteomics. In: Nordenfelt P, Collin M (eds) Bacterial pathogenesis. Methods in molecular biology, vol 1535. Humana Press, New York, NY, pp 17–24
Acknowledgments
The author thanks Lotta Happonen and Heiko Herwald (both Lund University, Lund, Sweden) for critical input.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Wilk, L. (2020). Protocol for Proteome Analysis of Group A Streptococcus. In: Proft, T., Loh, J. (eds) Group A Streptococcus. Methods in Molecular Biology, vol 2136. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0467-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0467-0_9
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0466-3
Online ISBN: 978-1-0716-0467-0
eBook Packages: Springer Protocols