Biomedical Research Center, National Institute on Aging, National Institutes of Health, 251 Bayview Boulevard, Suite 100, Baltimore, Maryland 21224-6825, USA.
Anal Chem. 2011 Oct 1;83(19):7400-7. doi: 10.1021/ac201403y. Epub 2011 Sep 2.
SIRT6 is a histone deacetylase that has been proposed as a potential therapeutic target for metabolic disorders and the prevention of age-associated diseases. Thus, the identification of compounds that modulate SIRT6 activity could be of great therapeutic importance. The aim of this study was to develop a screening method for the identification of novel modulators of SIRT6 from a natural plant extract. We immobilized SIRT6 onto the surface of magnetic beads, and assessed SIRT6 enzymatic activity on synthetic acetylated histone tails (H3K9Ac) by measuring products of the deacetylation process. The SIRT6 coated magnetic beads were then suspended in fenugreek seed extract (Trigonella foenum-graecum) as a bait to identify active ligands that suppress SIRT6 activity. While the entire extract also inhibited SIRT6 activity in a cell-based assay, the inhibitory effect of two flavonoids from this extract, quercetin and vitexin, was only detected in vitro. This is the first report on the use of protein-coated magnetic beads for the identification of an active ligand from a botanical matrix, and it sets the basis for the de novo identification of SIRT6 modulators from complex biological mixtures.
SIRT6 是一种组蛋白去乙酰化酶，被认为是治疗代谢紊乱和预防与年龄相关疾病的潜在治疗靶点。因此，鉴定能够调节 SIRT6 活性的化合物可能具有重要的治疗意义。本研究旨在从天然植物提取物中开发一种筛选 SIRT6 新型调节剂的方法。我们将 SIRT6 固定在磁珠表面，并通过测量去乙酰化过程的产物来评估 SIRT6 对合成乙酰化组蛋白尾（H3K9Ac）的酶活性。然后，将涂有 SIRT6 的磁珠悬浮在葫芦巴种子提取物（Trigonella foenum-graecum）中，以鉴定抑制 SIRT6 活性的有效配体。虽然整个提取物也在基于细胞的测定中抑制了 SIRT6 活性，但该提取物中的两种黄酮类化合物槲皮素和牡荆素仅在体外检测到抑制作用。这是首次报道使用蛋白质包被的磁珠从植物基质中鉴定活性配体，为从头鉴定复杂生物混合物中的 SIRT6 调节剂奠定了基础。
