4-Deoxy-4-fluoro-GalNAz (4FGalNAz) Is a Metabolic Chemical Reporter of O-GlcNAc Modifications, Highlighting the Notable Substrate Flexibility of O-GlcNAc Transferase.
4-Deoxy-4-fluoro-GalNAz (4FGalNAz) Is a Metabolic Chemical Reporter of O-GlcNAc Modifications, Highlighting the Notable Substrate Flexibility of O-GlcNAc Transferase.
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DOI：
10.1021/acschembio.1c00818
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发表时间：
2022-01-21
影响因子：
4
通讯作者：
Pratt MR
中科院分区：
文献类型：
--
作者：
Jackson EG;Cutolo G;Yang B;Yarravarapu N;Burns MWN;Bineva-Todd G;Roustan C;Thoden JB;Lin-Jones HM;van Kuppevelt TH;Holden HM;Schumann B;Kohler JJ;Woo CM;Pratt MR
生物正交化学已经彻底改变了许多领域。例如，糖基化的代谢化学报告物（MCR）是含有生物正交官能团的单糖的类似物，例如叠氮化物或炔。MCR通过生命系统代谢并入糖蛋白中，并且随后可以采用生物正交反应来安装可视化和富集标签。不幸的是，大多数MCR对一类糖基化没有选择性（例如，N-连锁与O-连锁），使可以收集的信息类型复杂化。我们和其他人通过改变MCR的结构并因此使其偏向于某些代谢途径和/或O-GlcNAc转移酶（OGT），成功创建了对细胞内O-GlcNAc修饰具有选择性的MCR。在这里，我们试图做同样的核心GalNAc残基的粘蛋白O-连接的糖基化。用于粘蛋白O-连接糖基化的最广泛应用的MCR GalNAz可以在4-羟基处酶促差向异构化以得到GlcNAz。这导致细胞表面和O-GlcNAc标记的混合物。我们推断，用氟取代GalNAz的4-羟基将锁定该位置的立体化学，使MCR更具选择性。合成后，我们发现4FGalNAz标记哺乳动物细胞中的多种蛋白质，并且不像4FGalNAc那样干扰内源性糖基化途径。然而，通过随后的蛋白质组学和生物化学表征，我们发现4FGalNAz并不广泛标记细胞表面糖蛋白，而是主要是OGT的底物。尽管这些结果有些出乎意料，但它们再次强调了OGT的巨大底物灵活性，这对一系列潜在的非生物和天然单糖对细胞内蛋白质的修饰具有有趣且重要的影响。
Bio-orthogonal chemistries have revolutionized many fields. For example, metabolic chemical reporters (MCRs) of glycosylation are analogues of monosaccharides that contain a bio-orthogonal functionality, such as azides or alkynes. MCRs are metabolically incorporated into glycoproteins by living systems, and bio-orthogonal reactions can be subsequently employed to install visualization and enrichment tags. Unfortunately, most MCRs are not selective for one class of glycosylation (e.g., N-linked vs O-linked), complicating the types of information that can be gleaned. We and others have successfully created MCRs that are selective for intracellular O-GlcNAc modification by altering the structure of the MCR and thus biasing it to certain metabolic pathways and/or O-GlcNAc transferase (OGT). Here, we attempt to do the same for the core GalNAc residue of mucin O-linked glycosylation. The most widely applied MCR for mucin O-linked glycosylation, GalNAz, can be enzymatically epimerized at the 4-hydroxyl to give GlcNAz. This results in a mixture of cell-surface and O-GlcNAc labeling. We reasoned that replacing the 4-hydroxyl of GalNAz with a fluorine would lock the stereochemistry of this position in place, causing the MCR to be more selective. After synthesis, we found that 4FGalNAz labels a variety of proteins in mammalian cells and does not perturb endogenous glycosylation pathways unlike 4FGalNAc. However, through subsequent proteomic and biochemical characterization, we found that 4FGalNAz does not widely label cell-surface glycoproteins but instead is primarily a substrate for OGT. Although these results are somewhat unexpected, they once again highlight the large substrate flexibility of OGT, with interesting and important implications for intracellular protein modification by a potential range of abiotic and native monosaccharides.
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影响因子：
15
作者：
Darabedian N;Gao J;Chuh KN;Woo CM;Pratt MR
通讯作者：
Pratt MR
代谢聚糖标记过程中全 O 乙酰化非天然单糖诱导的人工半胱氨酸 S-糖基化。
影响因子：
16.6
作者：
Qin, Wei;Qin, Ke;Chen, Xing
通讯作者：
Chen, Xing
影响因子：
4.8
作者：
Thoden, JB;Holden, HM
通讯作者：
Holden, HM
影响因子：
14.8
作者：
Ge Y;Ramirez DH;Yang B;D'Souza AK;Aonbangkhen C;Wong S;Woo CM
通讯作者：
Woo CM
DOI：
10.1073/pnas.1010045108
发表时间：
2011-02-22
影响因子：
11.1
作者：
Boyce, Michael;Carrico, Isaac S.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.