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Applications of diagonal chromatography for proteome‐wide characterization of protein modifications and activity‐based analyses
Author(s) -
Gevaert Kris,
Impens Francis,
Van Damme Petra,
Ghesquière Bart,
Hanoulle Xavier,
Vandekerckhove Joël
Publication year - 2007
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2007.06149.x
Subject(s) - proteome , chromatography , characterization (materials science) , chemistry , diagonal , computational biology , biology , materials science , biochemistry , nanotechnology , mathematics , geometry
Numerous gel‐free proteomics techniques have been reported over the past few years, introducing a move from proteins to peptides as bits of information in qualitative and quantitative proteome studies. Many shotgun proteomics techniques randomly sample thousands of peptides in a qualitative and quantitative manner but overlook the vast majority of protein modifications that are often crucial for proper protein structure and function. Peptide‐based proteomic approaches have thus been developed to profile a diverse set of modifications including, but not at all limited, to phosphorylation, glycosylation and ubiquitination. Typical here is that each modification needs a specific, tailor‐made analytical procedure. In this minireview, we discuss how one technique − diagonal reverse‐phase chromatography − is applied to study two different types of protein modification: protein processing and protein N‐glycosylation. Additionally, we discuss an activity‐based proteome study in which purine‐binding proteins were profiled by diagonal chromatography.