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一种用于定量蛋白质组学的新型氘代烷基化试剂。

A new deuterated alkylating agent for quantitative proteomics.

作者信息

Sebastiano Roberto, Citterio Attilio, Lapadula Marta, Righetti Pier Giorgio

机构信息

Department of Chemistry, Material and Engineering Chemistry, Politecnico of Milano, Via Mancinelli 7, Milano 20131, Italy.

出版信息

Rapid Commun Mass Spectrom. 2003;17(21):2380-6. doi: 10.1002/rcm.1206.

Abstract

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).

摘要

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)。

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