%0 Journal Article
%T Enrichment strategy of cysteine-containing peptides based on covalent chromatography<br>基于共价色谱分离的含半胱氨酸肽段富集策略
%A MI Wei
%A WANG Jing
%A YING Wantao
%A JIA Wei
%A CAI Yun
%A QIAN Xiaohong
%A <br>米薇
%A 王晶
%A 应万涛
%A 贾伟
%A 蔡耘
%A 钱小红
%J 色谱
%D 2010
%I 
%X Automated multidimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS) is routinely applied in large scale proteome profiling. However global proteome analysis remains a technical challenge due to the issues associated with sample complexity by tryptic digestion. The application of tag containing peptide enrichment approach for sample pre-separation could reduce the complexity of protein digest. Here, we demonstrated a simple and highly efficient cysteine-containing peptide enrichment method using a thiol specific covalent resin. The cysteine-containing peptides from the tryptic digests of the complex protein mixtures were selected by covalent chromatography based on thiol-disulfide exchange, identified by mass spectrometry. The strategy was firstly optimized and evaluated by using the tryptic peptides of bovine serum albumin (BSA). Then the method was applied with a relatively complicated sample from a five standard protein mixture. The results of these studies show that the enrichment method of cysteine-containing peptides is highly specific, efficient and reproducible. The effectiveness of this method in reducing the sample complexity and improving the identification of peptides by mass spectrometry has enabled high-throughput, automatic and large-scale qualitative and quantitative proteome analyses.
%K covalent chromatography
%K liquid chromatography-tandem mass spectrometry
%K cysteine-containing peptides
%K enrichment
%K proteome<br>共价色谱
%K 液相色谱-串联质谱
%K 含半胱氨酸肽
%K 富集
%K 蛋白质组
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=6E709DC38FA1D09A4B578DD0906875B5B44D4D294832BB8E&cid=6579068328FE643F&jid=4D81E042D77AFEC6881D14759692069C&aid=404DB53A41BB911A071247EC7D630EC3&yid=140ECF96957D60B2&vid=D3E34374A0D77D7F&iid=0B39A22176CE99FB&sid=6270DC1B5693DDAF&eid=DDD31293A7C7D057&journal_id=1000-8713&journal_name=色谱&referenced_num=0&reference_num=17