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Chemical Probing of the Human Sirtuin 5 Active Site Reveals Its Substrate Acyl Specificity and Peptide‐Based Inhibitors
Author(s) -
Roessler Claudia,
Nowak Theresa,
Pannek Martin,
Gertz Melanie,
Nguyen Giang T. T.,
Scharfe Michael,
Born Ilona,
Sippl Wolfgang,
Steegborn Clemens,
Schutkowski Mike
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201402679
Subject(s) - sirtuin , peptide , chemistry , nad+ kinase , biochemistry , active site , lysine , substrate (aquarium) , stereochemistry , acetylation , enzyme , biology , amino acid , gene , ecology
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.