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Enhanced transglycosylation activity of an Endo-F3 mutant by ligand-directed localization

文献类型：期刊论文

作者Zou, Xiangman1,2; Liu, Zhi1,3; Liu, Liya1; Shi, Wei1; Li, Wanzhen1,3; Guo, Zifen2; Tang, Feng1; Huang, Wei1,4,5
刊名ORGANIC & BIOMOLECULAR CHEMISTRY
出版日期2022-02-15
页码10
ISSN号1477-0520
DOI10.1039/d2ob00030j
通讯作者Guo, Zifen(guozifen76@163.com) ; Tang, Feng(tangfeng2013@simm.ac.cn) ; Huang, Wei(huangwei@simm.ac.cn)
英文摘要At present, numerous studies have been reported to remodel the N-glycans of therapeutic antibodies for the gain of functions. Among the ways of remodeling antibody N-glycans, the chemoenzymatic glycoengineering approach by endoglycosidase (ENGase) has been deeply investigated and provided a significant tool for IgG glycoengineering. Among these cases, the transglycosylation activity of Endo-F3, compared to Endo-S and S2, is insufficient and limits its power in remodeling IgG glycosylation. Herein, we chemically conjugated the Endo-F3 mutant D165A with an Fc binding peptide (FcBP), aiming to improve the affinity of Endo-F3 D165A to IgGs, and therefore enhance the transglycosylation activity of D165A. In this report, we investigated the conjugation site of FcBP to D165A and the linkers between them and found that the conjugation indeed significantly increases the transglycosylation activity of D165A. Meanwhile, we optimized the FcBP-D165A catalyzed transglycosylation process, including the enzyme quantity, oxazoline concentration, and so on. Finally, by this method, we remodeled the N-glycans of rituximab and trastuzumab into homogeneous S2G2F, G2F, GN2M3, and M3 types with decreased enzyme quantity, oxazoline ratio, and catalyzing time. This method not only provides an enhanced ENGase for IgG glycoengineering but also suggests that ligand-directed localization of enzymes is a potential strategy to enhance the activity of enzymes towards the targeted substrate.
WOS关键词CHEMOENZYMATIC SYNTHESIS ; STREPTOCOCCUS-PYOGENES ; THERAPEUTIC ANTIBODIES ; GLYCOSYNTHASE MUTANTS ; GLYCAN SPECIFICITY ; CRYSTAL-STRUCTURE ; IGG ANTIBODIES ; ENDOGLYCOSIDASE ; PROTEIN ; GLYCOSYLATION
资助项目National Natural Science Foundation of China (NSFC)[2187116] ; National Natural Science Foundation of China (NSFC)[82003574] ; Shanghai Municipal Science and Technology Major Project (the Shanghai Sail Program)[19YF1457100] ; Special Research Assistant Program (Chinese Academy of Sciences, CAS) ; National Key Research and Development Plan[2021YFE0200500] ; Key Project of the Education Department of Hunan Province[19A419]
WOS研究方向Chemistry
语种英语
WOS记录号WOS:000755234100001
出版者ROYAL SOC CHEMISTRY
源URL[http://119.78.100.183/handle/2S10ELR8/300424]  
专题中国科学院上海药物研究所
通讯作者Guo, Zifen; Tang, Feng; Huang, Wei
作者单位1.Chinese Acad Sci, CAS Ctr Excellence Mol Cell Sci, Ctr Biotherapeut Discovery Res, CAS Key Lab Receptor Res,Shanghai Inst Mat Med, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China
2.Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pharm & Pharmacol, Hunan Prov Key Lab Tumor Microenvironm Respons Dr, Hengyang 421001, Hunan, Peoples R China
3.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing 210023, Peoples R China
4.Univ Chinese Acad Sci, 19A Yuquan Rd, Beijing 100049, Peoples R China
5.Univ Chinese Acad Sci, Sch Pharmaceut Sci & Technol, Inst Adv Study, Hangzhou 310024, Peoples R China
推荐引用方式
GB/T 7714		 Zou, Xiangman,Liu, Zhi,Liu, Liya,et al. Enhanced transglycosylation activity of an Endo-F3 mutant by ligand-directed localization[J]. ORGANIC & BIOMOLECULAR CHEMISTRY,2022:10.
APA Zou, Xiangman.,Liu, Zhi.,Liu, Liya.,Shi, Wei.,Li, Wanzhen.,...&Huang, Wei.(2022).Enhanced transglycosylation activity of an Endo-F3 mutant by ligand-directed localization.ORGANIC & BIOMOLECULAR CHEMISTRY,10.
MLA Zou, Xiangman,et al."Enhanced transglycosylation activity of an Endo-F3 mutant by ligand-directed localization".ORGANIC & BIOMOLECULAR CHEMISTRY (2022):10.

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来源：上海药物研究所

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