Xue Sun, Lei Tao, Lin Yi, Yilan Ouyang, Naiyu Xu, Duxin Li, Robert J. Linhardt, Zhenqing Zhang. N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database[J]. Journal of Pharmaceutical Analysis, 2017, 7(2): 87-94.
Citation:
Xue Sun, Lei Tao, Lin Yi, Yilan Ouyang, Naiyu Xu, Duxin Li, Robert J. Linhardt, Zhenqing Zhang. N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database[J]. Journal of Pharmaceutical Analysis, 2017, 7(2): 87-94.
Xue Sun, Lei Tao, Lin Yi, Yilan Ouyang, Naiyu Xu, Duxin Li, Robert J. Linhardt, Zhenqing Zhang. N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database[J]. Journal of Pharmaceutical Analysis, 2017, 7(2): 87-94.
Citation:
Xue Sun, Lei Tao, Lin Yi, Yilan Ouyang, Naiyu Xu, Duxin Li, Robert J. Linhardt, Zhenqing Zhang. N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database[J]. Journal of Pharmaceutical Analysis, 2017, 7(2): 87-94.
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu 215021, China
Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 1108th Street, Troy, NY 12180, USA
Funds:
The authors appreciate Prof. Shiliang Wu (Soochow University) providing GC cell lines. The authors are grateful to the National Natural Science Foundation of China
Priority Academic Program Development of Jiangsu Higher Education Institutions
and the funding for Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases
The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. This work describes a strategy that combines an efficient release, labeling and liquid chromatography–mass spectral analysis with the use of a comprehensive database to analyze N-glycans. The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. N-glycans from four different well-studied glycoproteins were analyzed by this strategy. The results provided much more accurate and comprehensive data than that had been previously reported. This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences were compared corresponding to their differentiation states. Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis.