在完全中性 pH 条件下通过胶内胰蛋白酶消化验证蛋白质中二硫键的排布。

Verification of protein disulfide bond arrangement by in-gel tryptic digestion under entirely neutral pH conditions.

机构信息

Laboratory of Proteomic Sciences, 21st Century COE Program, Kyoto Pharmaceutical University, Kyoto, Japan.

出版信息

Proteomics. 2010 Apr;10(7):1505-9. doi: 10.1002/pmic.200900056.

DOI:10.1002/pmic.200900056

PMID:20127682

Abstract

To develop a concise proteomic procedure to verify the protein disulfide bond arrangement, non-reductive trypsin digestion of neuregulin 1-beta1 (176-246), a model disulfide-containing protein, was assessed by a proteolytic (18)O-labeling analysis. As a result, the commonly used in-gel tryptic digestion method has been improved for use entirely under neutral pH conditions. With this procedure, the disulfide arrangement of proteins could represent a clinical index candidate in pathological proteomic studies.

摘要

为了开发一种简洁的蛋白质组学方法来验证蛋白质中二硫键的排布，我们通过酶解（18）O 标记分析来评估神经调节蛋白 1-β1（176-246）（一种模型含二硫键的蛋白）的非还原型胰蛋白酶消化。结果，我们对常用的胶内胰蛋白酶消化方法进行了改进，使其完全在中性 pH 条件下进行。通过这种方法，蛋白质的二硫键排布可以作为病理蛋白质组学研究中的一个临床指标候选物。