A new deuterated alkylating agent for quantitative proteomics.
Weakly basic molecules containing a double bond, such as 2- and 4-vinylpyridine, are able to react and selectively alkylate -SH groups in proteins, thus preventing their re-oxidation to disulphide bridges. In contrast to conventional alkylating agents such as iodoacetamide and non-charged acrylamide derivatives, such molecules achieve 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 analyzing proteins in which the -SH groups should be blocked. Additionally, the use of vinylpyridines, partially or totally deuterated and thus with a mass difference compared with their non-deuterated counterparts of 4-7 Da, allows studies of induction/repression of protein synthesis (quantitative proteomics).
Department of Chemistry, Material and Engineering Chemistry, Politecnico of Milano, Via Mancinelli 7, Milano 20131, Italy.
Roberto
R
Sebastiano
Attilio
A
Citterio
Marta
M
Lapadula
Pier Giorgio
PG
Righetti
D000477:Alkylating Agents / Q000737:chemistry*; D000478:Alkylation; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003545:Cysteine / Q000032:analysis; D003903:Deuterium / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007553:Isotope Labeling; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D013501:Surface-Active Agents
D000477:Alkylating Agents; D011506:Proteins; D013501:Surface-Active Agents; D003903:Deuterium; D009113:Muramidase; D003545:Cysteine