title: pH sensing by FAK-His58 regulates focal adhesion remodeling.
abstract: Intracellular pH (pHi) dynamics regulates diverse cellular processes, including remodeling of focal adhesions. We now report that focal adhesion kinase (FAK), a key regulator of focal adhesion remodeling, is a pH sensor responding to physiological changes in pH. The initial step in FAK activation is autophosphorylation of Tyr397, which increased with higher pHi. We used a genetically encoded biosensor to show increased pH at focal adhesions as they mature during cell spreading. We also show that cells with reduced pHi had attenuated FAK-pY397 as well as defective cell spreading and focal adhesions. Mutagenesis studies indicated FAK-His58 is critical for pH sensing and molecular dynamics simulations suggested a model in which His58 deprotonation drives conformational changes that may modulate accessibility of Tyr397 for autophosphorylation. Expression of FAK-H58A in fibroblasts was sufficient to restore defective autophosphorylation and cell spreading at low pHi. These data are relevant to understanding cancer metastasis, which is dependent on increased pHi and FAK activity.
authors.affiliation: Department of Cell and Tissue Biology and 2 Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143.
authors.forename: Chang-Hoon
authors.initials: CH
authors.lastname: Choi
authors.forename: Bradley A
authors.initials: BA
authors.lastname: Webb
authors.forename: Michael S
authors.initials: MS
authors.lastname: Chimenti
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Jacobson
authors.forename: Diane L
authors.initials: DL
authors.lastname: Barber
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D027682:Cation Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D051417:Focal Adhesion Kinase 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D022001:Focal Adhesions / Q000502:physiology*; D006639:Histidine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D009154:Mutation / Q000235:genetics; D010766:Phosphorylation; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D000074082:Sodium-Hydrogen Exchanger 1; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D027682:Cation Transport Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C490997:Slc9a1 protein, mouse; D000074082:Sodium-Hydrogen Exchanger 1; D017923:Sodium-Hydrogen Exchangers; D006639:Histidine; D051417:Focal Adhesion Kinase 1; C496051:Ptk2 protein, mouse
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