title: A peptide N-terminal protection strategy for comprehensive glycoproteome analysis using hydrazide chemistry based method.
abstract: Enrichment of glycopeptides by hydrazide chemistry (HC) is a popular method for glycoproteomics analysis. However, possible side reactions of peptide backbones during the glycan oxidation in this method have not been comprehensively studied. Here, we developed a proteomics approach to locate such side reactions and found several types of the side reactions that could seriously compromise the performance of glycoproteomics analysis. Particularly, the HC method failed to identify N-terminal Ser/Thr glycopeptides because the oxidation of vicinal amino alcohol on these peptides generates aldehyde groups and after they are covalently coupled to HC beads, these peptides cannot be released by PNGase F for identification. To overcome this drawback, we apply a peptide N-terminal protection strategy in which primary amine groups on peptides are chemically blocked via dimethyl labeling, thus the vicinal amino alcohols on peptide N-termini are eliminated. Our results showed that this strategy successfully prevented the oxidation of peptide N-termini and significantly improved the coverage of glycoproteome.
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, 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, Dalian, China.
authors.forename: Hongqiang
authors.initials: H
authors.lastname: Qin
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Sun
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Huang
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Mao
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Cheng
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] Shanghai Key Laboratory of Functional Materials Chemistry, East China University of Science and Technology.
authors.forename: Hao
authors.initials: H
authors.lastname: Wan
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Yating
authors.initials: Y
authors.lastname: Yao
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, Dalian, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Dong
authors.affiliation: 1] CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China [2] University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhu
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, Dalian, China.
authors.forename: Fangjun
authors.initials: F
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, Dalian, 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, Dalian, China.
authors.forename: Hanfa
authors.initials: H
authors.lastname: Zou
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D030562:Databases, Protein; D054794:Glycomics / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry* / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis*; D006834:Hydrazines / Q000737:chemistry*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
chemical_list: D000596:Amino Acids; D006020:Glycopeptides; D006023:Glycoproteins; D006834:Hydrazines; D020543:Proteome; C029424:hydrazine