Department of Chemistry, University of Southern California, Los Angeles, California 90089, United States.
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390, United States.
ACS Chem Biol. 2021 Oct 15;16(10):1924-1929. doi: 10.1021/acschembio.1c00470. Epub 2021 Jul 20.
Metabolic chemical reports have fundamentally changed the way researchers study glycosylation. However, when administered as per--acetylated sugars, reporter molecules can participate in nonspecific chemical labeling of cysteine residues termed -glycosylation. Without detailed proteomic analyses, these labeling events can be indistinguishable from bona fide enzymatic labeling convoluting experimental results. Here, we report a solution in the synthesis and characterization of two reporter molecules functionalized at the anomeric position with hexanoic acid: 1-Hex-GlcNAlk and 1-Hex-6AzGlcNAc. Both reporters exhibit robust labeling over background with negligible amounts of nonspecific chemical labeling in cell lysates. This strategy serves as a template for the design of future reporter molecules allowing for more reliable interpretation of results.
代谢化学报告从根本上改变了研究人员研究糖基化的方式。然而，当作为乙酰化糖给药时，报告分子可以参与半胱氨酸残基的非特异性化学标记，称为 -糖基化。如果没有详细的蛋白质组学分析，这些标记事件与真正的酶标记相混淆，会使实验结果复杂化。在这里，我们报告了在糖醛位置上用己酸功能化的两种报告分子的合成和表征：1-己酰基-GlcNAc 和 1-己酰基-6 氮杂-GlcNAc。这两种报告分子在细胞裂解物中均表现出强烈的标记背景，且非特异性化学标记的量可忽略不计。该策略为设计未来的报告分子提供了模板，从而可以更可靠地解释结果。
