Process for the selective alkylation of -SH groups in proteins and peptides for the study of complex protein mixtures

摘要

Weakly basic molecules containing a double bond (in particular vinylpyridines) are able to react and selectively alkylate -SH groups in proteins, thus preventing their re-oxidation to disulphur bridges. Contrary to conventional alkylating agents, such as iodoacetamide, such molecules reach 100% alkylation of all -SH residues, even in complex proteins, without reacting with other functional groups. Their use is particularly effective in proteome analysis and more generally for analysing proteins in which the -SH groups should be blocked. Additionally, the use of vinylpyridines partially or totally deuterated, and thus with a mass difference as compared to non-deuterated vinylpyridines, allows studies of induction/repression of protein synthesis.