State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Reasearch, Nankai University, Haihe Education Park, 38 Tongyan Road, Tianjin 300353, China.
School of Basic Medical Sciences, Henan University Joint National Laboratory for Antibody Drug Engineering, Kaifeng, Henan 475004, China.
Bioorg Med Chem Lett. 2019 Mar 15;29(6):802-805. doi: 10.1016/j.bmcl.2019.01.021. Epub 2019 Jan 22.
O-Linked N-acetylglucosamine (O-GlcNAc) is an abundant posttranslationalmonosaccaride-modification found on Ser or Thr residues of intracellular proteins in most eukaryotes. The dynamic nature of O-GlcNAc has enabled researchers to modulate the stoichiometry of O-GlcNAc on proteins in order to investigate its function. Cell permeable small moleculars have proven invaluable tools to increase O-GlcNAc levels. Herein, using in vitro substrate screening, we identified GlcNAcF as an OGT-accepted but OGA-resistant sugar mimic. Cellular experiments with cell-permeable peracetylated-GlcNAcF (AcGlcNAcF) displayed that AcGlcNAcF was a potent tool to increase O-GlcNAc levels in several cell lines. Further, NIH3T3 cells interfered with OGT (siOGT) showed significant decreasing of O-GlcNAc levels with AcGlcNAcF treatment, indicating O-GlcNAcF was an OGT-dependent modification. In addition, cellular toxic assay confirmed O-GlcNAcF production has no significant effect on cell proliferation or viability. Thus, AcGlcNAcF represents a safe and dual regulator for both OGT and OGA, which will benefit the study of O-GlcNAc.
O-连接的 N-乙酰葡萄糖胺（O-GlcNAc）是一种丰富的、存在于真核生物细胞内蛋白丝氨酸或苏氨酸残基上的翻译后单糖修饰。O-GlcNAc 的动态性质使研究人员能够调节蛋白质上 O-GlcNAc 的化学计量，以研究其功能。细胞通透的小分子已被证明是增加 O-GlcNAc 水平的非常有价值的工具。在此，我们通过体外底物筛选，鉴定出 GlcNAcF 是一种 OGT 接受但 OGA 抗性的糖类似物。用细胞通透的乙酰化 GlcNAcF（AcGlcNAcF）进行细胞实验显示，AcGlcNAcF 是增加几种细胞系中 O-GlcNAc 水平的有效工具。此外，用干扰 OGT（siOGT）的 NIH3T3 细胞进行处理时，AcGlcNAcF 处理导致 O-GlcNAc 水平显著降低，表明 AcGlcNAcF 是一种 OGT 依赖性修饰。此外，细胞毒性测定证实 AcGlcNAcF 的产生对细胞增殖或活力没有显著影响。因此，AcGlcNAcF 代表了 OGT 和 OGA 的安全且双重调节剂，这将有益于 O-GlcNAc 的研究。
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