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N-Glycosidase Rudaea cellulosilytica (PNGase Rc) Column for the Characterization of Glycoprotein Conformational Epitopes by Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS)

文献类型：期刊论文

作者Deng, Ying5,6; Yang, Dongyuan5,6; Zheng, Wen4; Qi, Ao2,3,5; Yang, Yulin2,3,5; Shu, Zhulin2,3,5; Zhang, Fengyi2,3,5; Han, Hongbo2,5; Chen, Jin2,5; Zheng, Jie1,2,3,5,6
刊名ANALYTICAL LETTERS
出版日期2024-02-19
页码15
关键词Glycoprotein conformational epitope hydrogen/deuterium exchange mass spectrometry (HDX-MS) N-glycosidase (PNGase Rc) column Rudaea cellulosilytica
ISSN号0003-2719
DOI10.1080/00032719.2024.2320243
通讯作者Zheng, Jie(jzheng@simm.ac.cn)
英文摘要Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a sensitive technique used to study the conformational dynamics of proteins in solution. However, performing HDX-MS with N-linked glycoproteins is challenging. Analyzing glycoproteins through HDX-MS requires the use of N-glycoprotein deglycosylation enzyme (peptide: N-glycanase, abbreviated as PNGase) to deglycosylate glycoproteins. Traditional PNGases are inactive or have low activity under HDX-MS operating conditions. Here, we recommend the use of ultra-acidic N-glycopeptidase from PNGase Rc, which exhibits optimal enzymatic activity compared to other PNGases at pH 2.2. We covalently fixed PNGase Rc onto POROS resin for HDX-MS online deglycosylation. The results show that the immobilized PNGase Rc combined with pepsin column has good stability under various quenching conditions. Its online deglycosylation efficiency is better than other PNGases. Meanwhile, we used AlphaFold 2 and molecular dynamics (MD) to characterize the structure and stability of PNGase Rc and other PNGases, and demonstrated that PNGase Rc is the most stable.
WOS关键词PEPTIDE
资助项目National Natural Science Foundation of China
WOS研究方向Chemistry
语种英语
WOS记录号WOS:001179627300001
出版者TAYLOR & FRANCIS INC
源URL[http://119.78.100.183/handle/2S10ELR8/310160]  
专题中国科学院上海药物研究所
通讯作者Zheng, Jie
作者单位1.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing 210023, Jiangsu, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
3.Univ Chinese Acad Sci, Hangzhou Inst Adv Study, Sch Pharmaceut Sci & Technol, Hangzhou, Peoples R China
4.Hubei Yuanda Biotechnol, Wuhan, Peoples R China
5.Chinese Acad Sci, Shanghai Inst Mat Med, Drug Res Ctr Immunol Dis, Shanghai, Peoples R China
6.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing, Jiangsu, Peoples R China
推荐引用方式
GB/T 7714		 Deng, Ying,Yang, Dongyuan,Zheng, Wen,et al. N-Glycosidase Rudaea cellulosilytica (PNGase Rc) Column for the Characterization of Glycoprotein Conformational Epitopes by Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS)[J]. ANALYTICAL LETTERS,2024:15.
APA Deng, Ying.,Yang, Dongyuan.,Zheng, Wen.,Qi, Ao.,Yang, Yulin.,...&Zheng, Jie.(2024).N-Glycosidase Rudaea cellulosilytica (PNGase Rc) Column for the Characterization of Glycoprotein Conformational Epitopes by Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS).ANALYTICAL LETTERS,15.
MLA Deng, Ying,et al."N-Glycosidase Rudaea cellulosilytica (PNGase Rc) Column for the Characterization of Glycoprotein Conformational Epitopes by Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS)".ANALYTICAL LETTERS (2024):15.

入库方式： OAI收割

来源：上海药物研究所

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