通过气相色谱-质谱法对全甲基化肽段进行氨基酸序列测定。2-10纳摩尔水平化学衍生物形成的优化。
Amino acid sequence determination by g.l.c.--mass spectrometry of permethylated peptides. Optimization of the formation of chemical derivatives at the 2-10 nmol level.
作者信息
Rose K, Simona M G, Offord R E
出版信息
Biochem J. 1983 Nov 1;215(2):261-72. doi: 10.1042/bj2150261.
Abstract
A new technique is described that permits the permethylation of acylated peptides at the 2-10 nmol level. The presence of up to 400 micrograms of sodium dodecyl sulphate per sample does not affect the reaction yields. The technique, which is a miniaturization of the widely used methyl iodide/dimethylsulphinyl carbanion procedure, employs a layer of hexane to exclude moisture and oxygen from the reaction mixture. Analysis of the peptide derivatives by combined g.l.c.--mass spectrometry permits amino acid sequence information to be obtained. In addition to studies of digests of a model substrate (glucagon), the new permethylation technique has been used to identify a peptide of interest from a digest of a cytochrome and to define the N-termini of two proteins at the 5 nmol level.
摘要
本文描述了一种新技术，该技术可实现2 - 10纳摩尔水平的酰化肽全甲基化。每个样品中存在高达400微克的十二烷基硫酸钠不会影响反应产率。该技术是广泛使用的碘甲烷/二甲基亚砜基碳负离子方法的小型化，它采用一层己烷来排除反应混合物中的水分和氧气。通过气相色谱 - 质谱联用对肽衍生物进行分析可获得氨基酸序列信息。除了对模型底物（胰高血糖素）的消化产物进行研究外，这种新的全甲基化技术还被用于从细胞色素消化产物中鉴定出感兴趣的肽，并在5纳摩尔水平确定两种蛋白质的N端。
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