title: Crystal structures of human RIP2 kinase catalytic domain complexed with ATP-competitive inhibitors: Foundations for understanding inhibitor selectivity.
abstract: Receptor interacting protein 2 (RIP2) is an intracellular kinase and key signaling partner for the pattern recognition receptors NOD1 and NOD2 (nucleotide-binding oligomerization domain-containing proteins 1 and 2). As such, RIP2 represents an attractive target to probe the role of these pathways in disease. In an effort to design potent and selective inhibitors of RIP2 we established a crystallographic system and determined the structure of the RIP2 kinase domain in an apo form and also in complex with multiple inhibitors including AMP-PCP (β,γ-Methyleneadenosine 5'-triphosphate, a non-hydrolysable adenosine triphosphate mimic) and structurally diverse ATP competitive chemotypes identified via a high-throughput screening campaign. These structures represent the first set of diverse RIP2-inhibitor co-crystal structures and demonstrate that the protein possesses the ability to adopt multiple DFG-in as well as DFG-out and C-helix out conformations. These structures reveal key protein-inhibitor structural insights and serve as the foundation for establishing a robust structure-based drug design effort to identify both potent and highly selective inhibitors of RIP2 kinase.
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA. Electronic address: adam.k.charnley@gsk.com.
authors.forename: Adam K
authors.initials: AK
authors.lastname: Charnley
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK. Electronic address: maire.a.convery@gsk.com.
authors.forename: Máire A
authors.initials: MA
authors.lastname: Convery
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Ami
authors.initials: A
authors.lastname: Lakdawala Shah
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK.
authors.forename: Emma
authors.initials: E
authors.lastname: Jones
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK.
authors.forename: Philip
authors.initials: P
authors.lastname: Hardwicke
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK.
authors.forename: Angela
authors.initials: A
authors.lastname: Bridges
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Ouellette
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Totoritis
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Schwartz
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Bryan W
authors.initials: BW
authors.lastname: King
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: David D
authors.initials: DD
authors.lastname: Wisnoski
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: James
authors.initials: J
authors.lastname: Kang
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Eidam
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Bartholomew J
authors.initials: BJ
authors.lastname: Votta
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Gough
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Marquis
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: John
authors.initials: J
authors.lastname: Bertin
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapy Area, GlaxoSmithKline, 1250 S. Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Linda
authors.initials: L
authors.lastname: Casillas
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D015195:Drug Design; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D047428:Protein Kinase Inhibitors; D000255:Adenosine Triphosphate; C506253:RIPK2 protein, human; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2
keywords: CARD3; CARDIAK; Inhibitor selectivity; Kinase inhibitor; Kinase structure; RICK; RIP2; RIPK2; Structure-based drug design; X-ray cocrystal structure