title: Allosteric regulation of focal adhesion kinase by PIP₂ and ATP.
abstract: Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase that regulates cell signaling, proliferation, migration, and development. A major mechanism of regulation of FAK activity is an intramolecular autoinhibitory interaction between two of its domains--the catalytic and FERM domains. Upon cell adhesion to the extracellular matrix, FAK is being translocated toward focal adhesion sites and activated. Interactions of FAK with phosphoinositide phosphatidylinsositol-4,5-bis-phosphate (PIP₂) are required to activate FAK. However, the molecular mechanism of the activation remains poorly understood. Recent fluorescence resonance energy transfer experiments revealed a closure of the FERM-kinase interface upon ATP binding, which is reversed upon additional binding of PIP₂. Here, we addressed the allosteric regulation of FAK by performing all-atom molecular-dynamics simulations of a FAK fragment containing the catalytic and FERM domains, and comparing the dynamics in the absence or presence of ATP and PIP₂. As a major conformational change, we observe a closing and opening motion upon ATP and additional PIP₂ binding, respectively, in good agreement with the fluorescence resonance energy transfer experiments. To reveal how the binding of the regulatory PIP₂ to the FERM F2 lobe is transduced to the very distant F1/N-lobe interface, we employed force distribution analysis. We identified a network of mainly charged residue-residue interactions spanning from the PIP₂ binding site to the distant interface between the kinase and FERM domains, comprising candidate residues for mutagenesis to validate the predicted mechanism of FAK activation.
authors: Heidelberg Institute for Theoretical Studies, Heidelberg, Germany.
authors: Jing
authors: J
authors: Zhou
authors: Heidelberg Institute for Theoretical Studies, Heidelberg, Germany.
authors: Agnieszka
authors: A
authors: Bronowska
authors: Spanish National Cancer Research Centre, Madrid, Spain.
authors: Johanne
authors: J
authors: Le Coq
authors: Spanish National Cancer Research Centre, Madrid, Spain.
authors: Daniel
authors: D
authors: Lietha
authors: Heidelberg Institute for Theoretical Studies, Heidelberg, Germany; Interdisciplinary Center for Scientific Computing, Heidelberg, Germany. Electronic address: frauke.graeter@h-its.org.
authors: Frauke
authors: F
authors: Gräter
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000494:Allosteric Regulation / Q000187:drug effects; D000818:Animals; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*; D007295:Inositol Phosphates / Q000494:pharmacology*; D056004:Molecular Dynamics Simulation; D025341:Principal Component Analysis; D011487:Protein Conformation; D013816:Thermodynamics
chemical_list: D007295:Inositol Phosphates; D000255:Adenosine Triphosphate; C066454:inositol 4,5-bisphosphate; D051416:Focal Adhesion Protein-Tyrosine Kinases
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