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Sample Preparation and Processing for Quick, Universal, and Insightful Microbial Proteomics. | LitMetric

Sample Preparation and Processing for Quick, Universal, and Insightful Microbial Proteomics.

Methods Mol Biol

Département Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris-Saclay, CEA, INRAE, Bagnols-sur-Cèze, France.

Published: December 2024

AI Article Synopsis

Article Abstract

Next-generation shotgun proteomics is one of the most valuable tools for gaining insight into the function of organisms. By providing a list of peptides and abundance information, proteomics enables the identification of proteins, their quantities, posttranslational modifications, and localization. The most refined shotgun proteomics workflow involves protein extraction, trypsin digestion, ultrahigh-performance liquid chromatography coupled to high-resolution tandem mass spectrometry, and confident assignment of resulting spectra to peptide sequences. In this study, we present a versatile, time- and cost-efficient experimental workflow for protein extraction, digestion, and analysis that can be applied to any type of microorganism. Our experimental procedure exhibits superior sensitivity compared to gel-based protocols and can be used for comparative microbial proteomics to highlight key players that explain phenotypic differences between conditions or for proteotyping new microbial isolates for taxonomic purposes.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-4298-6_5DOI Listing

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