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Solid‐Phase Cross‐Linking (SPCL): A new tool for protein structure studies
Author(s) -
Paramelle David,
Enjalbal Christine,
Amblard Muriel,
Forest Eric,
Heymann Michaël,
Cantel Sonia,
Geourjon Christophe,
Martinez Jean,
Subra Gilles
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000029
Subject(s) - chemistry , mass spectrometry , peptide , reagent , covalent bond , combinatorial chemistry , chromatography , biochemistry , organic chemistry
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.