Department of Life Science, Laboratory of Functional Glycomics, Ajou University, San 5, Wonchon-dong, Suwon 443- 749, Korea.
Protein Pept Lett. 2022;29(9):769-774. doi: 10.2174/0929866529666220805151725.
Protein S-nitrosylation (SNO) and O-GlcNAcylation are important posttranslational modifications. The biological connection between SNO and O-GlcNAcylation is not clear.
We aim to identify the crosstalk between SNO and O-GlcNAcylation during heat-shock.
Ex vivo heat-shock on mouse tissues together with in vitro heat-shock on culture cells was performed and global levels of SNO and O-GlcNAcylation were analyzed with Biotin-switch assay (BSA) and RL2 immunoblots.
Heat-shock induces hypo-SNO in parallel with hyper-O-GlcNAcylation. Inverted induction of hypo-SNO and hyper-O-GlcNAcylation is globally progressed in a time-dependent manner.
Moreover, heat-shock ubiquitously facilitates S-denitrosylation (SdeNO) of endogenous SNO-proteins including SNO-OGT, SNO-Hsp70, SNO-Hsp90, SNO-Akt, and SNOactin. Particularly, SdeNO of SNO-OGT leads to enhanced OGT activity.
These findings provide mechanistic evidence that heat-shock triggers SdeNO of SNOOGT by which OGT activity is up-regulated, resulting in hyper-O-GlcNAcylation.
蛋白质 S-亚硝基化 (SNO) 和 O-GlcNAc 酰化是重要的翻译后修饰。SNO 和 O-GlcNAc 酰化之间的生物学联系尚不清楚。
我们旨在确定热休克过程中 SNO 和 O-GlcNAc 酰化之间的串扰。
对小鼠组织进行离体热休克和对培养细胞进行体外热休克，并通过生物素转换测定 (BSA) 和 RL2 免疫印迹分析来分析 SNO 和 O-GlcNAc 酰化的整体水平。
热休克诱导 SNO 水平降低，同时 O-GlcNAc 酰化水平升高。SNO 水平降低和 O-GlcNAc 酰化水平升高的反向诱导呈时间依赖性的全局进展。
此外，热休克普遍促进包括 SNO-OGT、SNO-Hsp70、SNO-Hsp90、SNO-Akt 和 SNO-肌动蛋白在内的内源性 SNO 蛋白的 S-去硝基化 (SdeNO)。特别是，SNO-OGT 的 SdeNO 导致 OGT 活性增强。
这些发现为热休克通过 SNO-OGT 的 SdeNO 触发 OGT 活性上调，导致 O-GlcNAc 酰化增加提供了机制证据。
