Galleano Iacopo, Schiedel Matthias, Jung Manfred, Madsen Andreas S, Olsen Christian A
Center for Biopharmaceuticals and Department of Drug Design & Pharmacology, University of Copenhagen , Universitetsparken 2, DK-2100, Copenhagen, Denmark.
Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg , Albertstraße 25, 79104 Freiburg im Breisgau, Germany.
J Med Chem. 2016 Feb 11;59(3):1021-31. doi: 10.1021/acs.jmedchem.5b01532. Epub 2016 Jan 20.
Sirtuins are important regulators of lysine acylation, which is implicated in cellular metabolism and transcriptional control. This makes the sirtuin class of enzymes interesting targets for development of small molecule probes with pharmaceutical potential. To achieve detailed profiling and kinetic insight regarding sirtuin inhibitors, it is important to have access to efficient assays. In this work, we report readily synthesized fluorogenic substrates enabling enzyme-economical evaluation of SIRT2 inhibitors in a continuous assay format as well as evaluation of the properties of SIRT2 as a long chain deacylase enzyme. Novel enzymatic activities of SIRT2 were thus established in vitro, which warrant further investigation, and two known inhibitors, suramin and SirReal2, were profiled against substrates containing ε-N-acyllysine modifications of varying length.
沉默调节蛋白是赖氨酸酰化的重要调节因子，而赖氨酸酰化与细胞代谢和转录控制有关。这使得沉默调节蛋白类酶成为开发具有药物潜力的小分子探针的有趣靶点。为了获得关于沉默调节蛋白抑制剂的详细图谱和动力学见解，获得高效的检测方法很重要。在这项工作中，我们报告了易于合成的荧光底物，能够以连续检测的形式对SIRT2抑制剂进行酶经济评估，并评估SIRT2作为长链脱酰酶的性质。因此，在体外确定了SIRT2的新酶活性，这值得进一步研究，并且针对含有不同长度ε-N-酰基赖氨酸修饰的底物对两种已知抑制剂苏拉明和SirReal2进行了分析。
