Sebastiano Roberto, Citterio Attilio, Lapadula Marta, Righetti Pier Giorgio
Department of Chemistry, Material and Engineering Chemistry, Politecnico of Milano, Via Mancinelli 7, Milano 20131, Italy.
Rapid Commun Mass Spectrom. 2003;17(21):2380-6. doi: 10.1002/rcm.1206.
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).
含有双键的弱碱性分子,如2-乙烯基吡啶和4-乙烯基吡啶,能够与蛋白质中的-SH基团发生反应并选择性地使其烷基化,从而防止它们重新氧化形成二硫键。与传统的烷基化剂如碘乙酰胺和不带电荷的丙烯酰胺衍生物不同,这类分子即使在复杂蛋白质中也能使所有-SH残基实现100%烷基化,而不会与其他官能团发生反应。它们在蛋白质组分析中特别有效,更广泛地用于分析需要封闭-SH基团的蛋白质。此外,使用部分或完全氘代的乙烯基吡啶,与未氘代的对应物相比,质量相差4-7 Da,可用于研究蛋白质合成的诱导/抑制(定量蛋白质组学)。