title: Improved single-step enrichment methods of cross-linked products for protein structure analysis and protein interaction mapping.
abstract: Chemical cross-linking/mass spectrometry (MS) is gradually developing into a routine method to investigate protein conformation and to decipher protein interaction networks. To increase identification rates of the frequently low abundant cross-linked products in LC/MS/MS experiments, fast and reliable sample preparation protocols are indispensable. We present simplified solid phase extraction methods using C18/SCX StageTips and mixed-mode OASIS MCX cartridges for a single-step enrichment of cross-linked products prior to LC/MS/MS analysis. Our improved protocols result in 3.5 to 4.6 times higher numbers of cross-link identifications for the model protein bovine serum albumin compared to non-processed samples.
authors.affiliation: Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-St. 4, 06120, Halle (Saale), Germany.
authors.forename: Rico
authors.initials: R
authors.lastname: Schmidt
authors.affiliation: Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-St. 4, 06120, Halle (Saale), Germany. andrea.sinz@pharmazie.uni-halle.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Sinz
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D011506:Proteins / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins
keywords: Cross-linking; Enrichment; Protein conformation; Protein-protein interactions; Solid phase extraction; Strong cation exchange