O-linked N-acetyl-β-d-glucosamine (O-GlcNAc) is a dynamic post-translational modification that modifies thousands of proteins. However, the roles and mechanisms of O-GlcNAcylation have been clarified in only a few proteins, and one of the main reasons for this is the lack of site-specific anti-O-GlcNAc antibodies. Recently, we found that SIRT1, which is an NAD⁺-dependent deacetylase, is O-GlcNAcylated at the serine 549 site (S549) and plays a cytoprotective role under stress. However, the mechanism underlying the roles of SIRT1 O-GlcNAcylation remains unclear. Here, we describe a site-specific antibody for SIRT1 O-GlcNAcylated at S549, named SIRT1-549-O. This antibody can be used for immunoprecipitation and western blotting assays, and it can be used to recognize the endogenous levels of both human and mouse SIRT1 O-GlcNAcylation. Therefore, this antibody not only provides an effective method to further understand the roles of SIRT1 O-GlcNAcylation but also makes it possible to discover the genetic and pharmacological factors that could regulate SIRT1 activity by modulating its O-GlcNAcylation.

中文翻译：

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SIRT1 O-GlcNAcy丝氨酸549位点特异性抗体的产生和表征

O-连接的N-乙酰基-β - d-葡萄糖胺（O-GlcNAc）是一种动态的翻译后修饰，可修饰成千上万种蛋白质。但是，O-GlcNAcy的作用和机理仅在少数几种蛋白质中得到了阐明，其主要原因之一是缺乏位点特异性的抗O-GlcNAc抗体。最近，我们发现SIRT1是NAD ⁺依赖的脱乙酰基酶，在丝氨酸549位点被S-OlclcNA化（S549），并在压力下起细胞保护作用。但是，SIRT1 O-GlcNAcylation作用的潜在机制尚不清楚。在这里，我们描述了SIRT1 O-GlcNAcylated在S549的位点特异性抗体，称为SIRT1-549-O。该抗体可以用于免疫沉淀和蛋白质印迹试验，并且可以用于识别人和小鼠SIRT1 O-GlcNAcylation的内源性水平。因此，该抗体不仅为进一步了解SIRT1 O-GlcNAcylation的作用提供了一种有效的方法，而且还可能发现可以通过调节其O-GlcNAcylation来调节SIRT1活性的遗传和药理因子。