SRT1720, SRT2183, SRT1460, and Resveratrol Are Not Direct Activators of SIRT1
SRT1720, SRT2183, SRT1460, and Resveratrol Are Not Direct Activators of SIRT1
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DOI：
10.1074/jbc.m109.088682
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发表时间：
2010-03-12
影响因子：
4.8
通讯作者：
Ahn, Kay
中科院分区：
文献类型：
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作者：
Pacholec, Michelle;Bleasdale, John E.;Ahn, Kay
Sirtuins 催化 NAD(+) 依赖性蛋白脱乙酰化，是转录、细胞凋亡、代谢和衰老的关键调节因子。人类有七种去乙酰化酶 (SIRT1-7)，SIRT1 被认为是热量限制下游途径的关键介质，已被证明可以延迟 2 型糖尿病等与年龄相关的疾病的发病并降低发病率。通过小鼠中 Sirt1 基因的转基因过表达或通过小分子激活剂白藜芦醇和 SRT1720 的药理激活来增加 SIRT1 活性，已在 2 型糖尿病啮齿动物模型中显示出有益的效果，表明 SIRT1 可能是一个有吸引力的治疗靶点。在此，我们通过利用天然底物（包括缺乏荧光团的 p53 衍生肽底物以及纯化的天然全长蛋白质底物 p53 和乙酰辅酶 A 合成酶 1）进行生化测定来评估所谓的 SIRT1 激活剂。 SRT1720、其结构相关化合物 SRT2183 和 SRT1460 以及白藜芦醇不会导致天然肽或全长蛋白质底物明显激活 SIRT1，但它们确实会使用含有共价连接荧光团的肽底物激活 SIRT1。利用核磁共振、表面等离子体共振和等温量热技术，我们提供了这些化合物直接与含荧光团的肽底物相互作用的证据。此外，我们证明 SRT1720 既不会降低高脂肪饮食小鼠的血浆葡萄糖，也不会提高线粒体能力。 SRT1720、SRT2183、SRT1460 和白藜芦醇对受体、酶、转运蛋白和离子通道表现出多种脱靶活性。综上所述，我们得出结论：SRT1720、SRT2183、SRT1460 和白藜芦醇不是 SIRT1 的直接激活剂。
Sirtuins catalyze NAD(+)-dependent protein deacetylation and are critical regulators of transcription, apoptosis, metabolism, and aging. There are seven human sirtuins (SIRT1-7), and SIRT1 has been implicated as a key mediator of the pathways downstream of calorie restriction that have been shown to delay the onset and reduce the incidence of age-related diseases such as type 2 diabetes. Increasing SIRT1 activity, either by transgenic overexpression of the Sirt1 gene in mice or by pharmacological activation by small molecule activators resveratrol and SRT1720, has shown beneficial effects in rodent models of type 2 diabetes, indicating that SIRT1 may represent an attractive therapeutic target. Herein, we have assessed purported SIRT1 activators by employing biochemical assays utilizing native substrates, including a p53-derived peptide substrate lacking a fluorophore as well as the purified native full-length protein substrates p53 and acetyl-CoA synthetase1. SRT1720, its structurally related compounds SRT2183 and SRT1460, and resveratrol do not lead to apparent activation of SIRT1 with native peptide or full-length protein substrates, whereas they do activate SIRT1 with peptide substrate containing a covalently attached fluorophore. Employing NMR, surface plasmon resonance, and isothermal calorimetry techniques, we provide evidence that these compounds directly interact with fluorophore-containing peptide substrates. Furthermore, we demonstrate that SRT1720 neither lowers plasma glucose nor improves mitochondrial capacity in mice fed a high fat diet. SRT1720, SRT2183, SRT1460, and resveratrol exhibit multiple off-target activities against receptors, enzymes, transporters, and ion channels. Taken together, we conclude that SRT1720, SRT2183, SRT1460, and resveratrol are not direct activators of SIRT1.