Specific Enrichment of Peptides with N-Terminal Serine/Threonine by a Solid-Phase Capture-Release Approach for Efficient Proteomics Analysis

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• 作者：Yating Yao ; Junfeng Huang ; Kai Cheng ; Yanbo Pan ; Hongqiang Qin ; Mingliang Ye ; Hanfa Zou
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：November 17, 2015
• 年：2015
• 卷：87
• 期：22
• 页码：11353-11360
• 全文大小：425K
• ISSN：1520-6882

文摘

A problem for 鈥渟hot-gun鈥?proteomics is that the peptides generated in the proteolysis step overwhelm the analytical capacity of current LC鈥揗S/MS systems. A straightforward approach to overcome this problem is to reduce the sample complexity by isolating the representative peptides of each protein. In this study, we presented a facile solid-phase capture-release approach to selectively enrich the peptides with N-terminal serine/threonine from protein digests. This method exploited the highly efficient reaction between an aldehyde group and a hydrazine group. The excellent performance of this approach was validated using synthetic peptides as well as complex protein digests. It was found that high enrichment specificity could be obtained and the identifications for complex samples with and without enrichment were highly complementary. Besides, the enrichment of peptides with serine/threonine adjacent to different protease cleavage sites demonstrated that our method was able to enrich peptides from protein digests in a sequence specific way. As a result, this new approach provides a simple way to reduce sample complexity and facilitates the identification of low-abundance proteins.