Department of Chemistry and Pharmacy, Medicinal Chemistry, Friedrich Alexander University Erlangen-Nürnberg, Nikolaus-Fiebiger-Strasse 10, 91058, Erlangen, Germany.
Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg, Albertstrasse 25, 79104, Freiburg im Breisgau, Germany.
Chembiochem. 2020 Apr 17;21(8):1161-1166. doi: 10.1002/cbic.201900527. Epub 2020 Jan 22.
We have discovered the sirtuin-rearranging ligands (SirReals) to be highly potent and selective inhibitors of the NAD -dependent lysine deacetylase Sirt2. Using a biotinylated SirReal in combination with biolayer interferometry, we previously observed a slow dissociation rate of the inhibitor-enzyme complex; this had been postulated to be the key to the high affinity and selectivity of SirReals. However, to attach biotin to the SirReal core, we introduced a triazole as a linking moiety; this was shown by X-ray co-crystallography to interact with Arg97 of the cofactor binding loop. Herein, we aim to elucidate whether the observed long residence time of the SirReals is induced mainly by triazole incorporation or is an inherent characteristic of the SirReal inhibitor core. We used the novel label-free switchSENSE® technology, which is based on electrically switchable DNA nanolevers, to prove that the long residence time of the SirReals is indeed caused by the core scaffold.
我们发现，组蛋白去乙酰化酶 Sirt2 的烟酰胺腺嘌呤二核苷酸（NAD）依赖性赖氨酸去乙酰化酶的 sirtuin 重排配体（SirReals）是高度有效和选择性的抑制剂。使用带有生物素的 SirReal 和生物层干涉测量法，我们之前观察到抑制剂-酶复合物的缓慢解离速率；这被认为是 SirReals 高亲和力和选择性的关键。然而，为了将生物素连接到 SirReal 核心上，我们引入了三唑作为连接部分；X 射线共晶结构显示该连接部分与辅因子结合环的 Arg97 相互作用。在此，我们旨在阐明观察到的 SirReals 的长停留时间主要是由三唑的掺入引起的，还是 SirReal 抑制剂核心的固有特征。我们使用了基于电可切换 DNA 纳米杆的新型无标记 switchSENSE®技术，证明了 SirReals 的长停留时间确实是由核心支架引起的。
