Supplementary material from "New chemical tools for probing activity and inhibition of the NAD+-dependent lysine deacylase sirtuin 2"
Version 2 2018-03-24, 06:53
Version 1 2018-03-08, 18:14
Posted on 2018-03-24 - 06:53
Sirtuins are NAD+-dependent protein deacylases capable of cleaving off acetyl as well as other acyl groups from the ɛ-amino group of lysines in histones and other substrate proteins. They have been reported as promising drug targets, and thus modulators of their activity are needed as molecular tools to uncover their biological function and as potential therapeutics. Here, we present new assay formats that complement existing assays for sirtuin biochemistry and cellular target engagement. Firstly, we report the development of a homogeneous fluorescence-based activity assay using unlabelled acylated peptides. Upon deacylation, the free lysine residue reacts with fluorescamine to form a fluorophore. Secondly, using click chemistry with a TAMRA-azide on a propargylated sirtuin inhibitor, we prepared the first fluorescently labelled small-molecule inhibitor of Sirt2. This is used in a binding assay, which is based on fluorescence polarization. We used it successfully to map potential inhibitor-binding sites and also to show cellular Sirt2 engagement. By means of these new assays, we were able to identify and characterize novel Sirt2 inhibitors out of a focused library screen. The binding of the identified Sirt2 inhibitors was rationalized by molecular docking studies. These new chemical tools thus can enhance further sirtuin research.This article is part of the discussion meeting issue ‘Frontiers in epigenetic chemical biology'.
CITE THIS COLLECTION
DataCite
3 Biotech
3D Printing in Medicine
3D Research
3D-Printed Materials and Systems
4OR
AAPG Bulletin
AAPS Open
AAPS PharmSciTech
Abhandlungen aus dem Mathematischen Seminar der Universität Hamburg
ABI Technik (German)
Academic Medicine
Academic Pediatrics
Academic Psychiatry
Academic Questions
Academy of Management Discoveries
Academy of Management Journal
Academy of Management Learning and Education
Academy of Management Perspectives
Academy of Management Proceedings
Academy of Management Review
Swyter, Sören; Schiedel, Matthias; Monaldi, Daria; Szunyogh, Sándor; Lehotzky, Attila; Rumpf, Tobias; et al. (2018). Supplementary material from "New chemical tools for probing activity and inhibition of the NAD+-dependent lysine deacylase sirtuin 2". The Royal Society. Collection. https://doi.org/10.6084/m9.figshare.c.4026274.v2
or
Select your citation style and then place your mouse over the citation text to select it.
SHARE
Usage metrics
Read the peer-reviewed publication
AUTHORS (9)
SS
Sören Swyter
MS
Matthias Schiedel
DM
Daria Monaldi
SS
Sándor Szunyogh
AL
Attila Lehotzky
TR
Tobias Rumpf
JO
Judit Ovádi
WS
Wolfgang Sippl
MJ
Manfred Jung