title: Protein tyrosine phosphorylation and the regulation of KCl cotransport in trout erythrocytes.
abstract: Electroneutral salt transporters are activated and deactivated by changes to the phosphorylation status either of the transporter itself or of other, as yet unidentified, regulatory proteins. We have studied the effects of an inhibitor of protein tyrosine kinase (PTK), genistein, upon KCl cotransport in trout erythrocytes. We show that Cl-dependent K fluxes activated by physiological stimuli, i.e. oxygenation and beta-adrenergic agonists, are rapidly and completely blocked by genistein, whilst the inactive analogue of genistein, daidzein, had no effect. By contrast, the protein tyrosine phosphatase (PTP) inhibitor, vanadate (V), caused a slow but strong activation of an inactive cotransporter. This vanadate (V) activated flux was inhibited by genistein as well as by the serine/threonine phosphatase (PSP) inhibitor, calyculin A. However, genistein had no effect upon the activation of the cotransporter by the protein (serine/threonine) kinase (PSK) inhibitor, staurosporine, or by N-ethylmaleimide, which also appears to act by inhibiting a PSK. These results are consistent with a sequential scheme of at least two tyrosine phosphorylation events which lie upstream to the serine/threonine phosphorylation sites in the signal transduction pathway leading from stimulus to transporter activation. The regulation of the activity of KCl cotransporter appears to involve a complex series of phosphorylation reactions.
authors.affiliation: Department of Environmental and Evolutionary Biology, University of Liverpool, PO Box 147, Liverpool L69 3BX, UK.
authors.forename: Y R
authors.initials: YR
authors.lastname: Weaver
authors.forename: A R
authors.initials: AR
authors.lastname: Cossins
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism*; D019833:Genistein; D007529:Isoflavones / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology; D008387:Marine Toxins; D017686:Oncorhynchus mykiss; D010080:Oxazoles / Q000494:pharmacology; D010766:Phosphorylation; D011189:Potassium Chloride / Q000097:blood*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D027981:Symporters; D014443:Tyrosine / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology; D000096922:K Cl- Cotransporters
chemical_list: D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D019833:Genistein; D007529:Isoflavones; D007545:Isoproterenol; D008387:Marine Toxins; D010080:Oxazoles; D011189:Potassium Chloride; D011505:Protein-Tyrosine Kinases; D019311:Staurosporine; D027981:Symporters; D014443:Tyrosine; D014638:Vanadates; D000096922:K Cl- Cotransporters; C059041:calyculin A