title: Amine Chemistry Method for Selective Enrichment of N-Linked Glycopeptides for Glycoproteomics Analysis.
abstract: An amine chemistry method was developed for the extraction of N-glycopeptides using amine-functionalized beads for glycoproteomics analysis. Two reductive amination reactions between primary amine and aldehyde were employed in this approach. The first one was to block the primary amines in the peptides by addition of formaldehyde and sodium cyanoborohydride into the peptide sample, and the second one was to couple the glycopeptides onto solid phase beads by incubating the glycopeptides containing aldehyde groups (oxidized by periodate) with the amine-functionalized beads in the presence of sodium cyanoborohydride. It was demonstrated that the blocking of primary amines in the peptides by the first reductive amination reaction prior to the periodate oxidation made the amine chemistry method very efficient and sensitive. This new method was validated by analysis of glycoprotein standards as well as proteome samples. It was found that this new method led to significant increase in the identification of N-glycosites compared with the conventional hydrazide chemistry method.
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.|University of Chinese Academy of Sciences , No. 19A Yuquan Road, Beijing 100049, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: The Second Affiliated Hospital of Dalian Medical University , 467 Zhongshan Road, Dalian 116027, China.
authors.forename: Deguang
authors.initials: D
authors.lastname: Sun
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.|Research Center for Drug Metabolism, School of Life Sciences, Jilin University , 2699 Qianjin Avenue, Changchun 130012, China.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Cong
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.|University of Chinese Academy of Sciences , No. 19A Yuquan Road, Beijing 100049, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Mao
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.|University of Chinese Academy of Sciences , No. 19A Yuquan Road, Beijing 100049, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Huang
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.|University of Chinese Academy of Sciences , No. 19A Yuquan Road, Beijing 100049, China.
authors.forename: Hongqiang
authors.initials: H
authors.lastname: Qin
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.|University of Chinese Academy of Sciences , No. 19A Yuquan Road, Beijing 100049, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Huang
authors.affiliation: The Second Affiliated Hospital of Dalian Medical University , 467 Zhongshan Road, Dalian 116027, China.
authors.forename: Liming
authors.initials: L
authors.lastname: Wang
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.
authors.forename: Mingliang
authors.initials: M
authors.lastname: Ye
authors.affiliation: CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.
authors.forename: Hanfa
authors.initials: H
authors.lastname: Zou
mesh_terms: D000588:Amines / Q000737:chemistry*; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D008113:Liver Neoplasms / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D000588:Amines; D006020:Glycopeptides
keywords: glycopeptide enrichment; glycoproteomics; reductive amination reaction