Practical considerations for improving the productivity of mass spectrometry-based proteomics.

Détails

ID Serval
serval:BIB_D8498C6B038D
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Practical considerations for improving the productivity of mass spectrometry-based proteomics.
Périodique
Chimia
Auteur(s)
Laskay U.A., Srzentić K., Fornelli L., Upir O., Kozhinov A.N., Monod M., Tsybin Y.O.
ISSN
0009-4293 (Print)
ISSN-L
0009-4293
Statut éditorial
Publié
Date de publication
2013
Peer-reviewed
Oui
Volume
67
Numéro
4
Pages
244-249
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Résumé
Mass spectrometry (MS) is currently the most sensitive and selective analytical technique for routine peptide and protein structure analysis. Top-down proteomics is based on tandem mass spectrometry (MS/ MS) of intact proteins, where multiply charged precursor ions are fragmented in the gas phase, typically by electron transfer or electron capture dissociation, to yield sequence-specific fragment ions. This approach is primarily used for the study of protein isoforms, including localization of post-translational modifications and identification of splice variants. Bottom-up proteomics is utilized for routine high-throughput protein identification and quantitation from complex biological samples. The proteins are first enzymatically digested into small (usually less than ca. 3 kDa) peptides, these are identified by MS or MS/MS, usually employing collisional activation techniques. To overcome the limitations of these approaches while combining their benefits, middle-down proteomics has recently emerged. Here, the proteins are digested into long (3-15 kDa) peptides via restricted proteolysis followed by the MS/MS analysis of the obtained digest. With advancements of high-resolution MS and allied techniques, routine implementation of the middle-down approach has been made possible. Herein, we present the liquid chromatography (LC)-MS/MS-based experimental design of our middle-down proteomic workflow coupled with post-LC supercharging.
Mots-clé
Aspartic Acid Endopeptidases/analysis, Bacterial Proteins/metabolism, Candida albicans/enzymology, Chromatography, Liquid/methods, Fungal Proteins/analysis, Peptide Fragments/analysis, Proteomics, Tandem Mass Spectrometry/methods
Pubmed
Web of science
Création de la notice
19/10/2014 9:32
Dernière modification de la notice
03/03/2018 21:50
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