title: Chemical probing of the human sirtuin 5 active site reveals its substrate acyl specificity and peptide-based inhibitors.
abstract: Sirtuins are NAD(+)-dependent deacetylases acting as sensors in metabolic pathways and stress response. In mammals there are seven isoforms. The mitochondrial sirtuin 5 is a weak deacetylase but a very efficient demalonylase and desuccinylase; however, its substrate acyl specificity has not been systematically analyzed. Herein, we investigated a carbamoyl phosphate synthetase 1 derived peptide substrate and modified the lysine side chain systematically to determine the acyl specificity of Sirt5. From that point we designed six potent peptide-based inhibitors that interact with the NAD(+) binding pocket. To characterize the interaction details causing the different substrate and inhibition properties we report several X-ray crystal structures of Sirt5 complexed with these peptides. Our results reveal the Sirt5 acyl selectivity and its molecular basis and enable the design of inhibitors for Sirt5.
authors.affiliation: Department of Enzymology, Institute of Biochemistry and Biotechnology, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120 Halle/Saale (Germany).
authors.forename: Claudia
authors.initials: C
authors.lastname: Roessler
authors.forename: Theresa
authors.initials: T
authors.lastname: Nowak
authors.forename: Martin
authors.initials: M
authors.lastname: Pannek
authors.forename: Melanie
authors.initials: M
authors.lastname: Gertz
authors.forename: Giang T T
authors.initials: GT
authors.lastname: Nguyen
authors.forename: Michael
authors.initials: M
authors.lastname: Scharfe
authors.forename: Ilona
authors.initials: I
authors.lastname: Born
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sippl
authors.forename: Clemens
authors.initials: C
authors.lastname: Steegborn
authors.forename: Mike
authors.initials: M
authors.lastname: Schutkowski
mesh_terms: D000215:Acylation; D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D015195:Drug Design; D006801:Humans; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation; D037761:Sirtuins / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D010455:Peptides; C508544:SIRT5 protein, human; D037761:Sirtuins
keywords: Sirt5; acylation; inhibitors; protein deacetylases; substrate specificity