Schultz Michael B, Rinaldi Conrad, Lu Yuancheng, Amorim João A, Sinclair David A
Department of Genetics, Blavatnik Institute, Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA, USA.
CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
Methods Mol Biol. 2019;1983:133-149. doi: 10.1007/978-1-4939-9434-2_8.
SIRT1 is an NAD-dependent lysine deacetylase that promotes healthy aging and longevity in diverse organisms. Small molecule allosteric activators of SIRT1 such as resveratrol and SRT2104 directly bind to the N-terminus of SIRT1 and lower the K for the protein substrate. In rodents, sirtuin-activating compounds (STACs) protect from age-related diseases and extend life span. In human clinical trials, STACs have a high safety profile and anti-inflammatory activities. Here, we describe methods for identifying and characterizing STACs, including production of recombinant protein, in vitro assays with recombinant protein, and cellular assays based on mitochondrial dynamics. The methods described in this chapter will facilitate this discovery of improved STACs, natural and synthetic, in the pursuit of interventions to treat age-related diseases.
SIRT1是一种依赖烟酰胺腺嘌呤二核苷酸（NAD）的赖氨酸脱乙酰酶，可促进多种生物体的健康衰老和延长寿命。SIRT1的小分子变构激活剂，如白藜芦醇和SRT2104，直接与SIRT1的N端结合，降低蛋白质底物的米氏常数（K）。在啮齿动物中，sirtuin激活化合物（STACs）可预防与年龄相关的疾病并延长寿命。在人体临床试验中，STACs具有高安全性和抗炎活性。在此，我们描述了鉴定和表征STACs的方法，包括重组蛋白的制备、重组蛋白的体外测定以及基于线粒体动力学的细胞测定。本章所述方法将有助于发现改良的天然和合成STACs，以寻求治疗与年龄相关疾病的干预措施。
