title: Specific Enrichment of Peptides with N-Terminal Serine/Threonine by a Solid-Phase Capture-Release Approach for Efficient Proteomics Analysis.
abstract: A problem for "shot-gun" proteomics is that the peptides generated in the proteolysis step overwhelm the analytical capacity of current LC-MS/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.
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.|University of Chinese Academy of Sciences , Beijing, 100049, China.
authors.forename: Yating
authors.initials: Y
authors.lastname: Yao
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.|University of Chinese Academy of Sciences , Beijing, 100049, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Huang
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.|University of Chinese Academy of Sciences , Beijing, 100049, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.|University of Chinese Academy of Sciences , Beijing, 100049, China.
authors.forename: Yanbo
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.
authors.forename: Hongqiang
authors.initials: H
authors.lastname: Qin
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.
authors.forename: Mingliang
authors.initials: M
authors.lastname: Ye
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS) , Dalian, 116023, China.
authors.forename: Hanfa
authors.initials: H
authors.lastname: Zou
mesh_terms: D002851:Chromatography, High Pressure Liquid; D006801:Humans; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012694:Serine / Q000737:chemistry* / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D013912:Threonine / Q000737:chemistry* / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D010455:Peptides; D013912:Threonine; D012694:Serine; D014357:Trypsin