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.
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HDAC11 has a narrow substrate specificity, in contrast to SIRT2, which cleaves these modifications with similar activity as Kmyr.
“This indicates a quite narrow substrate specificity in contrast to another long chain deacylase, SIRT2, which cleaves these modifications with similar activity as Kmyr (Galleano et al., 2016).”
From Histone deacetylase 11 is an ε-N-Myristoyllysine hydrolase (2018) · cites this paper as Galleano et al., 2016
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Histone deacetylase 11 is an ε-N-Myristoyllysine hydrolase
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- Authors
- Galleano, I., Schiedel, M., Jung, M., Madsen, A. S., & Olsen, C. A.
- Affiliations
- University of Copenhagen (DK)
- University of Freiburg (DE)
- Corresponding author
- Olsen, Christian A.
- Venue
- Journal of Medicinal Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2016-01-20
- Volume
- 59
- Issue
- 3
- Pages
- 1021-1031
- ISSN
- 0022-2623 (ISSN-L)
- 1520-4804
- OpenAlex
- W2319093261
- Cited by
- 53
- References
- 28
- Citations by year
- 2026: 3 · 2025: 5 · 2024: 3 · 2023: 6 · 2022: 3 · 2021: 6 · 2020: 3 · 2019: 6 · 2018: 9 · 2017: 8 · 2016: 1
- FWCI
- 4.59
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Medicinal Chemistry (publishedVersion)
- PubMed (publishedVersion)
- FreiDok plus (Universitätsbibliothek Freiburg) (submittedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (submittedVersion)
- Funders
- University of Copenhagen
- Teknologi og Produktion, Det Frie Forskningsråd (12-132328)
- European Cooperation in Science and Technology (CM1406)
- Lundbeckfonden
- Topics
- Health Sciences › Medicine › Geriatrics and Gerontology › Sirtuins and Resveratrol in Medicine
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Histone Deacetylase Inhibitors Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Keywords
- SIRT2, fluorogenic substrates, continuous assay, lysine acylation, SIRT2 inhibitors, suramin, enzyme kinetics, chemical probes
- MeSH terms
- Acetamides, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Enzyme Inhibitors, Fluorescent Dyes, Humans, Lysine, Models, Molecular, Structure-Activity Relationship, Substrate Specificity, Suramin, Thiazoles, Molecular Structure, Sirtuin 2
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