A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay: Substrate Screening and Inhibitor Evaluation

Iacopo Galleano; Matthias Schiedel; Manfred Jung; Andreas S Madsen; Christian A Olsen

doi:10.1021/acs.jmedchem.5b01532

A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay: Substrate Screening and Inhibitor Evaluation

Iacopo Galleano, Matthias Schiedel, Manfred Jung, Andreas S Madsen, Christian A Olsen

28 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	Journal of Medicinal Chemistry
Vol/bind	59
Udgave nummer	3
Sider (fra-til)	1021-31
Antal sider	11
ISSN	0022-2623
DOI	https://doi.org/10.1021/acs.jmedchem.5b01532
Status	Udgivet - 11 feb. 2016

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10.1021/acs.jmedchem.5b01532

Citationsformater

@article{27d0101a63a84aa287199dbf2b1e899a,

title = "A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay: Substrate Screening and Inhibitor Evaluation",

abstract = "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.",

keywords = "Acetamides, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Enzyme Inhibitors, Fluorescent Dyes, Humans, Lysine, Models, Molecular, Molecular Structure, Sirtuin 2, Structure-Activity Relationship, Substrate Specificity, Suramin, Thiazoles, Journal Article, Research Support, Non-U.S. Gov't",

author = "Iacopo Galleano and Matthias Schiedel and Manfred Jung and Madsen, {Andreas S} and Olsen, {Christian A}",

year = "2016",

month = feb,

day = "11",

doi = "10.1021/acs.jmedchem.5b01532",

language = "English",

volume = "59",

pages = "1021--31",

journal = "Journal of Medicinal Chemistry",

issn = "0022-2623",

publisher = "American Chemical Society",

number = "3",

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TY - JOUR

T1 - A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay

T2 - Substrate Screening and Inhibitor Evaluation

AU - Galleano, Iacopo

AU - Schiedel, Matthias

AU - Jung, Manfred

AU - Madsen, Andreas S

AU - Olsen, Christian A

PY - 2016/2/11

Y1 - 2016/2/11

N2 - 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.

AB - 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.

KW - Acetamides

KW - Dose-Response Relationship, Drug

KW - Drug Evaluation, Preclinical

KW - Enzyme Inhibitors

KW - Fluorescent Dyes

KW - Humans

KW - Lysine

KW - Models, Molecular

KW - Molecular Structure

KW - Sirtuin 2

KW - Structure-Activity Relationship

KW - Substrate Specificity

KW - Suramin

KW - Thiazoles

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1021/acs.jmedchem.5b01532

DO - 10.1021/acs.jmedchem.5b01532

M3 - Journal article

C2 - 26788965

SN - 0022-2623

VL - 59

SP - 1021

EP - 1031

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

IS - 3

ER -

A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay: Substrate Screening and Inhibitor Evaluation

Abstract

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