title: Structural mechanisms of allostery and autoinhibition in JNK family kinases.
abstract: c-Jun N-terminal (JNK) family kinases have a common peptide-docking site used by upstream activating kinases, substrates, scaffold proteins, and phosphatases, where the ensemble of bound proteins determines signaling output. Although there are many JNK structures, little is known about mechanisms of allosteric regulation between the catalytic and peptide-binding sites, and the activation loop, whose phosphorylation is required for catalytic activity. Here, we compare three structures of unliganded JNK3 bound to different peptides. These were compared as a class to structures that differ in binding of peptide, small molecule ligand, or conformation of the kinase activation loop. Peptide binding induced an inhibitory interlobe conformer that was reversed by alterations in the activation loop. Structure class analysis revealed the subtle structural mechanisms for allosteric signaling between the peptide-binding site and activation loop. Biochemical data from isothermal calorimetry, fluorescence energy transfer, and enzyme inhibition demonstrated affinity differences among the three peptides that were consistent with structural observations.
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, 130 Scripps Way, Jupiter, FL 33458, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Laughlin
authors.forename: Jerome C
authors.initials: JC
authors.lastname: Nwachukwu
authors.forename: Mariana
authors.initials: M
authors.lastname: Figuera-Losada
authors.forename: Lisa
authors.initials: L
authors.lastname: Cherry
authors.forename: Kendall W
authors.initials: KW
authors.lastname: Nettles
authors.forename: Philip V
authors.initials: PV
authors.lastname: LoGrasso
mesh_terms: D051698:Activating Transcription Factor 2 / Q000737:chemistry; D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry; D000494:Allosteric Regulation; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006801:Humans; D048057:Mitogen-Activated Protein Kinase 10 / Q000037:antagonists & inhibitors / Q000737:chemistry*; D048055:Mitogen-Activated Protein Kinase 8 / Q000737:chemistry; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry; D017433:Protein Structure, Secondary; D013379:Substrate Specificity
chemical_list: C497105:ATF2 protein, human; D051698:Activating Transcription Factor 2; D048868:Adaptor Proteins, Signal Transducing; C490666:MAPK8IP1 protein, human; D009842:Oligopeptides; D047428:Protein Kinase Inhibitors; C112230:SH3BP5 protein, human; D048057:Mitogen-Activated Protein Kinase 10; D048055:Mitogen-Activated Protein Kinase 8