Chapter title |
Enrichment of Cell Surface-Associated Proteins in Gram-Positive Bacteria by Biotinylation or Trypsin Shaving for Mass Spectrometry Analysis
|
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Chapter number | 4 |
Book title |
Microbial Proteomics
|
Published in |
Methods in molecular biology, September 2018
|
DOI | 10.1007/978-1-4939-8695-8_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8693-4, 978-1-4939-8695-8
|
Authors |
Florian Bonn, Sandra Maaß, Jan Maarten van Dijl, Bonn, Florian, Maaß, Sandra, van Dijl, Jan Maarten |
Abstract |
In microbial cells surface-exposed proteins represent a physiologically important class of molecules as they enable these cells to interact with their environment both as free-living organisms and during interactions with a host. However, the characteristics of these proteins are quite divergent, which makes attempts to enrich, analyze, and quantify these molecules a challenging task. In this chapter two complementary methods for the enrichment and identification of cell surface-associated proteins, namely the biotinylation and the shaving approaches, are presented. Both protocols have been optimized for Gram-positive bacteria, and we provide a step-by-step guide for sample generation. Possible pitfalls during protein preparation are discussed. |
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