title: Insights into Lysine Deacetylation of Natively Folded Substrate Proteins by Sirtuins.
abstract: Sirtuins are NAD(+)-dependent lysine deacylases, regulating a variety of cellular processes. The nuclear Sirt1, the cytosolic Sirt2, and the mitochondrial Sirt3 are robust deacetylases, whereas the other sirtuins have preferences for longer acyl chains. Most previous studies investigated sirtuin-catalyzed deacylation on peptide substrates only. We used the genetic code expansion concept to produce natively folded, site-specific, and lysine-acetylated Sirt1-3 substrate proteins, namely Ras-related nuclear, p53, PEPCK1, superoxide dismutase, cyclophilin D, and Hsp10, and analyzed the deacetylation reaction. Some acetylated proteins such as Ras-related nuclear, p53, and Hsp10 were robustly deacetylated by Sirt1-3. However, other reported sirtuin substrate proteins such as cyclophilin D, superoxide dismutase, and PEPCK1 were not deacetylated. Using a structural and functional approach, we describe the ability of Sirt1-3 to deacetylate two adjacent acetylated lysine residues. The dynamics of this process have implications for the lifetime of acetyl modifications on di-lysine acetylation sites and thus constitute a new mechanism for the regulation of proteins by acetylation. Our studies support that, besides the primary sequence context, the protein structure is a major determinant of sirtuin substrate specificity.
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and.
authors.forename: Philipp
authors.initials: P
authors.lastname: Knyphausen
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and.
authors.forename: Susanne
authors.initials: S
authors.lastname: de Boor
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and.
authors.forename: Nora
authors.initials: N
authors.lastname: Kuhlmann
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and.
authors.forename: Lukas
authors.initials: L
authors.lastname: Scislowski
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and.
authors.forename: Antje
authors.initials: A
authors.lastname: Extra
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and.
authors.forename: Linda
authors.initials: L
authors.lastname: Baldus
authors.affiliation: the Institute for Biochemistry, Zülpicher Strasse 47b, University of Cologne, 50674 Cologne, Germany.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Schacherl
authors.affiliation: the Institute for Biochemistry, Zülpicher Strasse 47b, University of Cologne, 50674 Cologne, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Baumann
authors.affiliation: the Institute for Biochemistry, Zülpicher Strasse 47b, University of Cologne, 50674 Cologne, Germany.
authors.forename: Ines
authors.initials: I
authors.lastname: Neundorf
authors.affiliation: From the Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Strasse 26, University of Cologne, 50931 Cologne and michael.lammers@uni-koeln.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Lammers
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D002151:Calorimetry; D003460:Crystallization; D008239:Lysine / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017510:Protein Folding; D037761:Sirtuins / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D037761:Sirtuins; D008239:Lysine
keywords: GTPase; acetylation; p53; sirtuin; synthetic biology