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| An in vitro assay for enzymatic studies on human ALG13/14 heterodimeric UDP-N-acetylglucosamine transferase | |
| Wang, Chun-Di1; Xu, Si1; Chen, Shuai1; Chen, Zheng-Hui1; Dean, Neta2; Wang, Ning1; Gao, Xiao-Dong1,3 | |
| 2022-09-19 | |
| Source Publication | FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
 |
| ISSN | 2296-634X |
| Volume | 10Pages:15 |
| Abstract | The second step of eukaryotic lipid-linked oligosaccharide (LLO) biosynthesis is catalyzed by the conserved ALG13/ALG14 heterodimeric UDP-N-acetylglucosamine transferase (GnTase). In humans, mutations in ALG13 or ALG14 lead to severe neurological disorders with a multisystem phenotype, known as ALG13/14-CDG (congenital disorders of glycosylation). How these mutations relate to disease is unknown because to date, a reliable GnTase assay for studying the ALG13/14 complex is lacking. Here we describe the development of a liquid chromatography/mass spectrometry-based quantitative GnTase assay using chemically synthesized GlcNAc-pyrophosphate-dolichol as the acceptor and purified human ALG13/14 dimeric enzyme. This assay enabled us to demonstrate that in contrast to the literature, only the shorter human ALG13 isoform 2, but not the longer isoform 1 forms a functional complex with ALG14 that participates in LLO synthesis. The longer ALG13 isoform 1 does not form a complex with ALG14 and therefore lacks GnTase activity. Importantly, we further established a quantitative assay for GnTase activities of ALG13- and ALG14-CDG variant alleles, demonstrating that GnTase deficiency is the cause of ALG13/14-CDG phenotypes. |
| Keyword | N-glycosylation lipid-linked oligosaccharide (LLO) ALG glycosyltransferases ALG13/14 UDP-N-acetylglucosamine transferase ALG13 isoforms congenital disorders of glycosylation (CDG) |
| DOI | 10.3389/fcell.2022.1008078 |
| Language | 英语 |
| WOS Keyword | CONGENITAL DISORDERS ; GLYCOSYLATION ; ALG2 ; OLIGOSACCHARIDES ; 2ND-STEP ; SEIZURES ; SPECTRUM ; VARIANT ; FORM |
| Funding Project | National Natural Science Foundation of China[21807048] ; National Natural Science Foundation of China[31971216] ; National Natural Science Foundation of China[22077053] ; Jiangsu Planned Projects for Postdoctoral Research Funds[2020Z167] ; Shandong Provincial Major Scientific and Technological Innovation Project[2019JZZY011006] ; Top-notch Academic Programs Project of Jiangsu Higher Education Institutions ; Program of Introducing Talents of Discipline to Universities[111-2-06] ; Special fund for Zaozhuang Excellence agglomeration project ; Qing Lan Project of Jiangsu Province |
| WOS Research Area | Cell Biology ; Developmental Biology |
| WOS Subject | Cell Biology ; Developmental Biology |
| Funding Organization | National Natural Science Foundation of China ; Jiangsu Planned Projects for Postdoctoral Research Funds ; Shandong Provincial Major Scientific and Technological Innovation Project ; Top-notch Academic Programs Project of Jiangsu Higher Education Institutions ; Program of Introducing Talents of Discipline to Universities ; Special fund for Zaozhuang Excellence agglomeration project ; Qing Lan Project of Jiangsu Province |
| WOS ID | WOS:000863482900001 |
| Publisher | FRONTIERS MEDIA SA |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ipe.ac.cn/handle/122111/54798 |
| Collection | 中国科学院过程工程研究所 |
| Corresponding Author | Wang, Ning; Gao, Xiao-Dong |
| Affiliation | 1.Jiangnan Univ, Sch Biotechnol, Key Lab Carbohydrate Chem & Biotechnol, Minist Educ, Wuxi, Peoples R China 2.SUNY Stony Brook, Dept Biochem & Cell Biol, New York, NY USA 3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing, Peoples R China |
| Recommended Citation GB/T 7714 | Wang, Chun-Di,Xu, Si,Chen, Shuai,et al. An in vitro assay for enzymatic studies on human ALG13/14 heterodimeric UDP-N-acetylglucosamine transferase[J]. FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY,2022,10:15. |
| APA | Wang, Chun-Di.,Xu, Si.,Chen, Shuai.,Chen, Zheng-Hui.,Dean, Neta.,...&Gao, Xiao-Dong.(2022).An in vitro assay for enzymatic studies on human ALG13/14 heterodimeric UDP-N-acetylglucosamine transferase.FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY,10,15. |
| MLA | Wang, Chun-Di,et al."An in vitro assay for enzymatic studies on human ALG13/14 heterodimeric UDP-N-acetylglucosamine transferase".FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY 10(2022):15. |
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