title: Crystallographic structure of a small molecule SIRT1 activator-enzyme complex.
abstract: SIRT1, the founding member of the mammalian family of seven NAD(+)-dependent sirtuins, is composed of 747 amino acids forming a catalytic domain and extended N- and C-terminal regions. We report the design and characterization of an engineered human SIRT1 construct (mini-hSIRT1) containing the minimal structural elements required for lysine deacetylation and catalytic activation by small molecule sirtuin-activating compounds (STACs). Using this construct, we solved the crystal structure of a mini-hSIRT1-STAC complex, which revealed the STAC-binding site within the N-terminal domain of hSIRT1. Together with hydrogen-deuterium exchange mass spectrometry (HDX-MS) and site-directed mutagenesis using full-length hSIRT1, these data establish a specific STAC-binding site and identify key intermolecular interactions with hSIRT1. The determination of the interface governing the binding of STACs with human SIRT1 facilitates greater understanding of STAC activation of this enzyme, which holds significant promise as a therapeutic target for multiple human diseases.
authors.affiliation: 1] Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA [2] GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Han
authors.identifier: 0000000210048142
authors.initials: H
authors.lastname: Dai
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: April W
authors.initials: AW
authors.lastname: Case
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Thomas V
authors.initials: TV
authors.lastname: Riera
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Considine
authors.affiliation: ExSAR Corporation, 11 Deer Park Drive, Suite 103, Monmouth Junction, New Jersey 08852, USA.
authors.forename: Jessica E
authors.initials: JE
authors.lastname: Lee
authors.affiliation: ExSAR Corporation, 11 Deer Park Drive, Suite 103, Monmouth Junction, New Jersey 08852, USA.
authors.forename: Yoshitomo
authors.initials: Y
authors.lastname: Hamuro
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Huizhen
authors.identifier: 0000000267428656
authors.initials: H
authors.lastname: Zhao
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Sweitzer
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Beth
authors.initials: B
authors.lastname: Pietrak
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Schwartz
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Blum
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Jeremy S
authors.initials: JS
authors.lastname: Disch
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Caldwell
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Szczepankiewicz
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Oalmann
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Pui
authors.initials: P
authors.lastname: Yee Ng
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Brian H
authors.initials: BH
authors.lastname: White
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Casaubon
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Radha
authors.initials: R
authors.lastname: Narayan
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Karsten
authors.identifier: 000000030789015X
authors.initials: K
authors.lastname: Koppetsch
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Francis
authors.initials: F
authors.lastname: Bourbonais
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, 350 Shushanhu Road, Hefei, Anhui Province 230031, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Wu
authors.affiliation: High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Sciences, 350 Shushanhu Road, Hefei, Anhui Province 230031, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Viva Biotech, 334 Aidisheng Road, Zhangjiang High-tech Park, Shanghai 201203, China.
authors.forename: Dongming
authors.initials: D
authors.lastname: Qian
authors.affiliation: Viva Biotech, 334 Aidisheng Road, Zhangjiang High-tech Park, Shanghai 201203, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Jiang
authors.affiliation: Viva Biotech, 334 Aidisheng Road, Zhangjiang High-tech Park, Shanghai 201203, China.
authors.forename: Cheney
authors.initials: C
authors.lastname: Mao
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Minghui
authors.initials: M
authors.lastname: Wang
authors.affiliation: GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: Erding
authors.initials: E
authors.lastname: Hu
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Joe C
authors.initials: JC
authors.lastname: Wu
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: Robert B
authors.initials: RB
authors.lastname: Perni
authors.affiliation: Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.
authors.forename: George P
authors.initials: GP
authors.lastname: Vlasuk
authors.affiliation: 1] Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA [2] GlaxoSmithKline, 1250S. Collegeville Road, Collegeville, Pennsylvania 19426, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Ellis
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D003460:Crystallization; D018360:Crystallography, X-Ray; D041961:Deuterium Exchange Measurement; D004926:Escherichia coli; D005822:Genetic Vectors; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D013058:Mass Spectrometry; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D056564:Sirtuin 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: C447939:SIRT1 protein, human; D056564:Sirtuin 1; D008239:Lysine