title: Studying SIRT6 regulation using H3K56 based substrate and small molecules.
abstract: SIRT6 is a modulator of chromatin structure having an important role in healthy ageing, and there is a crucial need to find specific modulators for it. Therefore, the activity of SIRT6 should be studied using a variety of methods. We examined the capability of SIRT6 to deacetylate a set of five fluorogenic substrates based on p53 and histone H3 sequences. The substrate designed around H3K56 deacetylation site exhibited the best signal-to-background ratio and was chosen for further studies. Nicotinamide is a known inhibitor for sirtuins, and it was found to be less potent inhibitor for SIRT6 than it is for SIRT1. In addition, we studied 15 other small molecule sirtuin modulators using the H3K56 based substrate. EX-527, quercetin and three pseudopeptidic compounds were found to be the most potent SIRT6 inhibitors, exhibiting over 50% deacetylation inhibition. These findings describe the first modulators of SIRT6 activity at the physiologically important H3K56 deacetylation site.
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211 Kuopio, Finland.
authors.forename: Piia
authors.initials: P
authors.lastname: Kokkonen
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211 Kuopio, Finland. Electronic address: Minna.Rahnasto@uef.fi.
authors.forename: Minna
authors.initials: M
authors.lastname: Rahnasto-Rilla
authors.affiliation: Istituto Pasteur - Fondazione Cenci Bolognetti, Dipartimento di Chimica e Tecnologie del Farmaco, Università degli Studi di Roma "La Sapienza", P.le A. Moro 5, 00185 Rome, Italy. Electronic address: paolo.mellini@uniroma1.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Mellini
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211 Kuopio, Finland.
authors.forename: Elina
authors.initials: E
authors.lastname: Jarho
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211 Kuopio, Finland.
authors.forename: Maija
authors.initials: M
authors.lastname: Lahtela-Kakkonen
authors.affiliation: School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211 Kuopio, Finland. Electronic address: Tarja.Kokkola@uef.fi.
authors.forename: Tarja
authors.initials: T
authors.lastname: Kokkola
mesh_terms: D000107:Acetylation / Q000187:drug effects; D002227:Carbazoles / Q000737:chemistry / Q000494:pharmacology; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015394:Molecular Structure; D009536:Niacinamide / Q000737:chemistry / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D011794:Quercetin / Q000737:chemistry / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D037761:Sirtuins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: C550547:6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide; D002227:Carbazoles; D006657:Histones; D010455:Peptides; D011994:Recombinant Proteins; D009536:Niacinamide; D011794:Quercetin; C516592:SIRT6 protein, human; D037761:Sirtuins
keywords: Fluorometric assay; Histone deacetylase; Nicotinamide; SIRT6; Sirtuins