title: Evidence for activation of BK Ca channels by a known inhibitor of focal adhesion kinase, PF573228.
abstract: OBJECTIVE
  PF573228 is an inhibitor of focal adhesion kinase (FAK) and recognized to affect cell adhesion and migration in many types of cells. Its effects on ion currents and membrane potential have been investigated in this study.
  
  
  METHODS
  Electrophysiological studies of PF573228 actions on ion currents were performed in pituitary tumor (GH(3)) cells, in GH(3) cells transfected with K(Ca)1.1 siRNAs and in human embryonic kidney (HEK) cells expressing α-human slowpoke (α-hSlo).
  
  
  RESULTS
  In whole-cell experiments, PF573228 reversibly increased the amplitude of Ca(2+)-activated K(+) currents (I(K(Ca))) in GH(3) cells. In inside-out recordings, this compound added to the bath did not modify single-channel conductance but stimulated large-conductance Ca(2+)-activated K(+) (BK(Ca)) channels with an EC(50) value of 3.2 μM. As BK(Ca)-channel activity was stimulated by PF573228 (3 μM), subsequent application of BMS191011 (3 μM) did not further increase channel activity. PF573228 shifted the activation curve of BK(Ca) channels to less positive membrane potential. Change in the kinetic behavior of BK(Ca) channels caused by this compound is a result of the increased backward rate constants between closed states. PF573228 depressed the firing of action potentials in GH(3) cells. However, in GH(3) cells transfected with K(Ca)1.1 siRNAs, PF573228-stimulated I(K(Ca)) was abolished. In HEK293T cells expressing α-hSlo, PF573228 enhanced BK(Ca)-channel activity.
  
  
  CONCLUSIONS
  In addition to an inhibition of FAK phosphorylation, PF573228 is effective in activating BK(Ca) channels. The direct stimulation of these channels by this compound may contribute to the underlying mechanism through which it influences cell behavior.
authors.affiliation: Department of Anaesthesiology, Chi-Mei Medical Center, Tainan City, Taiwan.
authors.forename: Edmund Cheung
authors.initials: EC
authors.lastname: So
authors.forename: King Chuen
authors.initials: KC
authors.lastname: Wu
authors.forename: Chia-Hua
authors.initials: CH
authors.lastname: Liang
authors.forename: Jen-Yin
authors.initials: JY
authors.lastname: Chen
authors.forename: Sheng-Nan
authors.initials: SN
authors.lastname: Wu
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D045744:Cell Line, Tumor; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D057809:HEK293 Cells; D006801:Humans; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / Q000235:genetics; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000187:drug effects* / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D010911:Pituitary Neoplasms / Q000473:pathology; D015363:Quinolones / Q000494:pharmacology*; D051381:Rats; D013450:Sulfones / Q000494:pharmacology*; D014162:Transfection
chemical_list: C521108:6-(4-(3-(methylsulfonyl)benzylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydroquinolin-2(1H)-one; C494841:KCNMA1 protein, human; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D051036:Large-Conductance Calcium-Activated Potassium Channels; D015363:Quinolones; D013450:Sulfones; D051416:Focal Adhesion Protein-Tyrosine Kinases