title: Human kidney anion exchanger 1 localisation in MDCK cells is controlled by the phosphorylation status of two critical tyrosines.
abstract: An important question in renal physiology is how the alpha-intercalated cells of the kidney regulate the distribution of the basolateral kidney anion exchanger 1 (kAE1) according to systemic acid-base status. Previous work using a MDCKI model system demonstrated that kAE1 basolateral targeting requires an N-terminal determinant and a critical C-terminal tyrosine (Y904). Here, we show that the N-terminal determinant is residue Y359, because a Y359A substitution mutant was mistargeted to the apical membrane. Further determinants might exist because a range of N-terminal kAE1 truncations that contained Y359 were incorrectly targeted to the TGN. Y359 and Y904 in kAE1 are phosphorylated upon pervanadate treatment and this phosphorylation is sensitive to specific Src kinase family inhibitors. We tested a range of stimuli on this model system and only the application of high nonphysiological concentrations of extracellular bicarbonate, and to a lesser extent hypertonicity or hyperosmolarity, induced tyrosine phosphorylation of kAE1. Treatment with pervanadate caused internalisation of kAE1 from the plasma membrane, but treatment with high concentrations of bicarbonate did not, because of the hypertonicity of the solution. We propose that alpha-intercalated cells control the distribution of kAE1 by reversible phosphorylation of tyrosine residues Y359 and Y904.
authors.affiliation: University of Bristol, Department of Biochemistry, School of Medical Sciences, University Walk, Bristol BS8 1TD, UK.
authors.forename: Rosalind C
authors.initials: RC
authors.lastname: Williamson
authors.forename: Alice C N
authors.initials: AC
authors.lastname: Brown
authors.forename: William J
authors.initials: WJ
authors.lastname: Mawby
authors.forename: Ashley M
authors.initials: AM
authors.lastname: Toye
mesh_terms: D000136:Acid-Base Equilibrium / Q000502:physiology*; D019943:Amino Acid Substitution; D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000235:genetics / Q000378:metabolism*; D001639:Bicarbonates / Q000494:pharmacology; D002021:Buffers; D002460:Cell Line; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D020125:Mutation, Missense; D010766:Phosphorylation / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D001639:Bicarbonates; D002021:Buffers; D004791:Enzyme Inhibitors; C076715:pervanadate; D014638:Vanadates; D014443:Tyrosine