Another turn of the screw in shaving Gram-positive bacteria: Optimization of proteomics surface protein identification in Streptococcus pneumoniae

Journal of Proteomics
Alfonso Olaya-AbrilManuel J Rodríguez-Ortega

Abstract

Bacterial surface proteins are of outmost importance as they play critical roles in the interaction between cells and their environment. In addition, they can be targets of either vaccines or antibodies. Proteomic analysis through "shaving" live cells with proteases has become a successful approach for a fast and reliable identification of surface proteins. However, this protocol has not been able to reach the goal of excluding cytoplasmic contamination, as cell lysis is an inherent process during culture and experimental manipulation. In this work, we carried out the optimization of the "shaving" strategy for the Gram-positive human pathogen Streptococcus pneumoniae, a bacterium highly susceptible to autolysis, and set up the conditions for maximizing the identification of surface proteins containing sorting or exporting signals, and for minimizing cytoplasmic contamination. We also demonstrate that cell lysis is an inherent process during culture and experimental manipulation, and that a low level of lysis is enough to contaminate a "surfome" preparation with peptides derived from cytoplasmic proteins. When the optimized conditions were applied to several clinical isolates, we found the majority of the proteins described to i...Continue Reading

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Citations

Jan 7, 2014·Journal of Proteome Research·Thomas PribylSven Hammerschmidt
Apr 30, 2013·Journal of Proteomics·Alfonso Olaya-AbrilManuel J Rodríguez-Ortega
Apr 29, 2014·Journal of Proteomics·Alfonso Olaya-AbrilManuel J Rodríguez-Ortega
Jul 15, 2015·Proteomics. Clinical Applications·Allison C Galassie, Andrew J Link
Jul 31, 2012·Journal of Proteomics·Lidia Gómez-GascónManuel J Rodríguez-Ortega
May 15, 2015·Analytical and Bioanalytical Chemistry·Javier Alfonso-GarridoJose L Luque-Garcia
Aug 2, 2016·Comparative Immunology, Microbiology and Infectious Diseases·Lidia Gómez-GascónInmaculada Luque
Jul 18, 2015·Molecular & Cellular Proteomics : MCP·Alfonso Olaya-AbrilManuel J Rodríguez-Ortega
Nov 21, 2018·Omics : a Journal of Integrative Biology·Julia EsbelinMichel Hébraud
Jun 6, 2018·Frontiers in Microbiology·Elvira MarínConcha Gil
Jun 20, 2018·PeerJ·Irene Jiménez-MunguíaManuel J Rodríguez-Ortega
Jun 5, 2017·Scientific Reports·Watcharapong MitsuwanManuel J Rodríguez-Ortega
May 10, 2019·Frontiers in Nutrition·Rosa Anna SicilianoMaria Fiorella Mazzeo

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