title: Solid-Phase Cross-Linking (SPCL): a new tool for protein structure studies.
abstract: A wide range of chemical reagents are available to study the protein-protein interactions or protein structures. After reaction with such chemicals, covalently modified proteins are digested, resulting in shorter peptides that are analyzed by mass spectrometry (MS). Used especially when NMR of X-ray data are lacking, this methodology requires the identification of modified species carrying relevant information, among the unmodified peptides. To overcome the drawbacks of existing methods, we propose a more direct strategy relying on the synthesis of solid-supported cleavable monofunctional reagents and cross-linkers that react with proteins and that selectively release, after protein digestion and washings, the modified peptide fragments ready for MS analysis. Using this Solid-Phase Cross-Linking (SPCL) strategy, only modified sequences are analyzed and consistent data can be easily obtained since the signals of interest are not masked or suppressed by over-represented unmodified materials.
authors.affiliation: Institut des Biomolécules Max Mousseron, CNRS, Universités Montpellier 1 et 2, Montpellier, France.
authors.forename: David
authors.initials: D
authors.lastname: Paramelle
authors.forename: Christine
authors.initials: C
authors.lastname: Enjalbal
authors.forename: Muriel
authors.initials: M
authors.lastname: Amblard
authors.forename: Eric
authors.initials: E
authors.lastname: Forest
authors.forename: Michaël
authors.initials: M
authors.lastname: Heymann
authors.forename: Sonia
authors.initials: S
authors.lastname: Cantel
authors.forename: Christophe
authors.initials: C
authors.lastname: Geourjon
authors.forename: Jean
authors.initials: J
authors.lastname: Martinez
authors.forename: Gilles
authors.initials: G
authors.lastname: Subra
mesh_terms: D000818:Animals; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000737:chemistry*; D006736:Horses; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014357:Trypsin / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D010446:Peptide Fragments; D011506:Proteins; D014357:Trypsin