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Substrates and Cyclic Peptide Inhibitors of the Oligonucleotide-Activated Sirtuin 7** |
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| Authors: | Julie E. Bolding Alexander L. Nielsen Iben Jensen Tobias N. Hansen Line A. Ryberg Samuel T. Jameson Pernille Harris Günther H. J. Peters John M. Denu Joseph M. Rogers Christian A. Olsen |
| Affiliation: | 1. Center for Biopharmaceuticals & Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark These authors contributed equally to this work.;2. Center for Biopharmaceuticals & Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Jagtvej 160, 2100 Copenhagen, Denmark;3. Department of Chemistry, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark Current address: Department of Immunology and Microbiology, University of Copenhagen, 2200 Copenhagen, Denmark;4. Department of Chemistry, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark Current address: Department of Chemistry, University of Copenhagen, 2200 Copenhagen, Denmark;5. Department of Chemistry, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark;6. Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI 53706 USA Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715 USA |
| Abstract: | The sirtuins are NAD+-dependent lysine deacylases, comprising seven isoforms (SIRT1–7) in humans, which are involved in the regulation of a plethora of biological processes, including gene expression and metabolism. The sirtuins share a common hydrolytic mechanism but display preferences for different ϵ-N-acyllysine substrates. SIRT7 deacetylates targets in nuclei and nucleoli but remains one of the lesser studied of the seven isoforms, in part due to a lack of chemical tools to specifically probe SIRT7 activity. Here we expressed SIRT7 and, using small-angle X-ray scattering, reveal SIRT7 to be a monomeric enzyme with a low degree of globular flexibility in solution. We developed a fluorogenic assay for investigation of the substrate preferences of SIRT7 and to evaluate compounds that modulate its activity. We report several mechanism-based SIRT7 inhibitors as well as de novo cyclic peptide inhibitors selected from mRNA-display library screening that exhibit selectivity for SIRT7 over other sirtuin isoforms, stabilize SIRT7 in cells, and cause an increase in the acetylation of H3 K18. |
| Keywords: | Epigenetics Histones Posttranslational Modification Lysine acylation mRNA Display |