School of Chemistry and Biochemistry and the Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, 30332, USA.
Angew Chem Int Ed Engl. 2017 Jun 12;56(25):7107-7111. doi: 10.1002/anie.201702191. Epub 2017 May 17.
Glycoproteins contain a wealth of valuable information regarding the development and disease status of cells. In cancer cells, some glycans (such as the Tn antigen) are highly up-regulated, but this remains largely unknown for glycoproteins with a particular glycan. Herein, an innovative method combining enzymatic and chemical reactions was first designed to enrich glycoproteins with the Tn antigen. Using synthetic glycopeptides with O-GalNAc (the Tn antigen) or O-GlcNAc, we demonstrated that the method is selective for glycopeptides with O-GalNAc and can distinguish between these two modifications. The diagnostic ions from the tagged O-GalNAc further confirmed the effectiveness of the method and confidence in the identification of glycopeptides with the Tn antigen by mass spectrometry. Using this method, we identified 96 glycoproteins with the Tn antigen in Jurkat cells. The method can be extensively applied in biological and biomedical research.
糖蛋白包含了大量关于细胞发育和疾病状态的有价值的信息。在癌细胞中，一些聚糖（如 Tn 抗原）被高度上调，但对于具有特定聚糖的糖蛋白，这在很大程度上仍然未知。在此，我们首次设计了一种结合酶和化学反应的创新方法来富集 Tn 抗原的糖蛋白。使用带有 O-GalNAc（Tn 抗原）或 O-GlcNAc 的合成糖肽，我们证明该方法对带有 O-GalNAc 的糖肽具有选择性，并且可以区分这两种修饰。标记的 O-GalNAc 的诊断离子进一步证实了该方法的有效性，并对通过质谱法鉴定 Tn 抗原糖肽的方法有了信心。使用这种方法，我们在 Jurkat 细胞中鉴定出了 96 种带有 Tn 抗原的糖蛋白。该方法可以广泛应用于生物和生物医学研究。
