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Chemical Probing of the Human Sirtuin 5 Active Site Reveals Its Substrate Acyl Specificity and Peptide‐Based Inhibitors |
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| Authors: | Claudia Roessler Theresa Nowak Martin Pannek Dr Melanie Gertz Giang T T Nguyen Michael Scharfe Dr Ilona Born Prof?Dr Wolfgang Sippl Prof?Dr Clemens Steegborn Prof?Dr Mike Schutkowski |
| Affiliation: | 1. Department of Enzymology, Institute of Biochemistry and Biotechnology, Martin‐Luther‐University Halle‐Wittenberg, Kurt‐Mothes‐Strasse 3, 06120 Halle/Saale (Germany);2. Department of Medical Chemistry, Institute of Pharmacy, Martin‐Luther‐University Halle‐Wittenberg, Halle/Saale (Germany);3. Department of Biochemistry, University of Bayreuth (Germany) |
| 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. |
| Keywords: | acylation inhibitors protein deacetylases Sirt5 substrate specificity |
