| Autor: |
Juyoux P; European Molecular Biology Laboratory (EMBL), Grenoble, France., Galdadas I; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva, Switzerland.; School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland., Gobbo D; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva, Switzerland.; School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland., von Velsen J; European Molecular Biology Laboratory (EMBL), Grenoble, France., Pelosse M; European Molecular Biology Laboratory (EMBL), Grenoble, France., Tully M; European Synchrotron Radiation Facility, Grenoble, France., Vadas O; Protein and peptide purification platform, Faculty of Medicine, University of Geneva, Geneva, Switzerland., Gervasio FL; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva, Switzerland.; School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland.; Department of Chemistry, University College London, London, UK.; Institute of Structural and Molecular Biology, University College London, London, UK.; Swiss Institute of Bioinformatics, Geneva, Switzerland., Pellegrini E; European Molecular Biology Laboratory (EMBL), Grenoble, France., Bowler MW; European Molecular Biology Laboratory (EMBL), Grenoble, France. |
| Abstrakt: |
The mitogen-activated protein kinase (MAPK) p38α is a central component of signaling in inflammation and the immune response and is, therefore, an important drug target. Little is known about the molecular mechanism of its activation by double phosphorylation from MAPK kinases (MAP2Ks), because of the challenge of trapping a transient and dynamic heterokinase complex. We applied a multidisciplinary approach to generate a structural model of p38α in complex with its MAP2K, MKK6, and to understand the activation mechanism. Integrating cryo-electron microscopy with molecular dynamics simulations, hydrogen-deuterium exchange mass spectrometry, and experiments in cells, we demonstrate a dynamic, multistep phosphorylation mechanism, identify catalytically relevant interactions, and show that MAP2K-disordered amino termini determine pathway specificity. Our work captures a fundamental step of cell signaling: a kinase phosphorylating its downstream target kinase. |
