Systematic identification of the protein substrates of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase-T1/T2/T3 using a human proteome microarray
Systematic identification of the protein substrates of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase-T1/T2/T3 using a human proteome microarray
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使用人类蛋白质组微阵列系统鉴定 UDP-GalNAc：多肽 N-乙酰半乳糖胺基转移酶-T1/T2/T3 的蛋白质底物
DOI：
10.1002/pmic.201600485
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发表时间：
2017
期刊：
影响因子：
3.4
通讯作者：
Zhang Yan
中科院分区：
文献类型：
--
作者：
Xu Zhijue;Li Xing;Zhou Shumin;Xie Wenxian;Wang Jing;Cheng Li;Wang Sheng;Guo Shujuan;Xu Zhaowei;Cao Xin;Zhang Menghui;Yu Biao;Narimatsu Hisashi;Tao Sheng-Ce;Zhang Yan
O - GalNAc糖基化是粘蛋白O -型糖基化的第一步。在人类中，它是由20个同源的UDP‐GalNAc：多肽N‐乙酰半乳糖氨基转移酶（ppGalNAc‐Ts）家族催化的。到目前为止，关于它们的蛋白质底物特异性的信息非常有限。在这项研究中，我们开发了一种芯片上的ppGalNAc‐Ts检测方法，可以快速系统地识别每个ppGalNAc‐T的蛋白质底物。详细地说，我们利用人类蛋白质组微阵列作为蛋白质底物，UDP‐GalNAz作为核苷酸糖供体进行点击化学检测。从总共16368个人类蛋白中，我们鉴定出570个ppGalNAc‐T1、T2和T3的潜在底物。其中128个底物重叠，其余为同工异构体特异性。进一步的聚类分析表明，与ppGalNAc‐T2相比，ppGalNAc‐T1与ppGalNAc‐T3具有更密切的系统发育关系，这与这些ppGalNAc‐Ts系统发育树的拓扑结构一致。综上所述，我们基于微阵列的酶促分析全面揭示了ppGalNAc‐T1、T2和T3的底物特征，这不仅为它们的部分功能冗余提供了合理的解释，而且清楚地暗示了每种酶在不同生物过程中的一些特殊作用。
O‐GalNAc glycosylation is the initial step of the mucin‐typeO‐glycosylation. In humans, it is catalyzed by a family of 20 homologous UDP‐GalNAc:polypeptide N‐acetylgalactosaminyltransferases (ppGalNAc‐Ts). So far, there is very limited information on their protein substrate specificities. In this study, we developed an on‐chip ppGalNAc‐Ts assay that could rapidly and systematically identify the protein substrates of each ppGalNAc‐T. In detail, we utilized a human proteome microarray as the protein substrates and UDP‐GalNAz as the nucleotide sugar donor for click chemistry detection. From a total of 16 368 human proteins, we identified 570 potential substrates of ppGalNAc‐T1, T2, and T3. Among them, 128 substrates were overlapped, while the rest were isoform specific. Further cluster analysis of these substrates showed that the substrates of ppGalNAc‐T1 had a closer phylogenetic relationship with that of ppGalNAc‐T3 compared with ppGalNAc‐T2, which was consistent with the topology of the phylogenetic tree of these ppGalNAc‐Ts. Taken together, our microarray‐based enzymatic assay comprehensively reveals the substrate profile of the ppGalNAc‐T1, T2, and T3, which not only provides a plausible explanation for their partial functional redundancy as reported, but clearly implies some specialized roles of each enzyme in different biological processes.