title: Applications of diagonal chromatography for proteome-wide characterization of protein modifications and activity-based analyses.
abstract: 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.
authors.affiliation: Department of Medical Protein Research, VIB, Ghent, Belgium. kris.gevaert@ugent.be
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
authors.forename: Francis
authors.initials: F
authors.lastname: Impens
authors.forename: Petra
authors.initials: P
authors.lastname: Van Damme
authors.forename: Bart
authors.initials: B
authors.lastname: Ghesquière
authors.forename: Xavier
authors.initials: X
authors.lastname: Hanoulle
authors.forename: Joël
authors.initials: J
authors.lastname: Vandekerckhove
mesh_terms: D000595:Amino Acid Sequence; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D011506:Proteins; D020543:Proteome