title: Mouse Ae1 E699Q mediates SO42-i/anion-o exchange with [SO42-]i-dependent reversal of wild-type pHo sensitivity.
abstract: The SLC4A1/AE1 gene encodes the electroneutral Cl(-)/HCO(3)(-) exchanger of erythrocytes and renal type A intercalated cells. AE1 mutations cause familial spherocytic and stomatocytic anemias, ovalocytosis, and distal renal tubular acidosis. The mutant mouse Ae1 polypeptide E699Q expressed in Xenopus oocytes cannot mediate Cl(-)/HCO(3)(-) exchange or (36)Cl(-) efflux but exhibits enhanced dual sulfate efflux mechanisms: electroneutral exchange of intracellular sulfate for extracellular sulfate (SO(4)(2-)(i)/SO(4)(2-)(o) exchange), and electrogenic exchange of intracellular sulfate for extracellular chloride (SO(4)(2-)(i)/Cl(-)(o) exchange). Whereas wild-type AE1 mediates 1:1 H(+)/SO(4)(2-) cotransport in exchange for either Cl(-) or for the H(+)/SO(4)(2-) ion pair, mutant Ae1 E699Q transports sulfate without cotransport of protons, similar to human erythrocyte AE1 in which the corresponding E681 carboxylate has been chemically converted to the alcohol (hAE1 E681OH). We now show that in contrast to the normal cis-stimulation by protons of wild-type AE1-mediated SO(4)(2-) transport, both SO(4)(2-)(i)/Cl(-)(o) exchange and SO(4)(2-)(i)/SO(4)(2-)(o) exchange mediated by mutant Ae1 E699Q are inhibited by acidic pH(o) and activated by alkaline pH(o). hAE1 E681OH displays a similarly altered pH(o) dependence of SO(4)(2-)(i)/Cl(-)(o) exchange. Elevated [SO(4)(2-)](i) increases the K(1/2) of Ae1 E699Q for both extracellular Cl(-) and SO(4)(2-), while reducing inhibition of both exchange mechanisms by acid pH(o). The E699Q mutation also leads to increased potency of self-inhibition by extracellular SO(4)(2-). Study of the Ae1 E699Q mutation has revealed the existence of a novel pH-regulatory site of the Ae1 polypeptide and should continue to provide valuable paths toward understanding substrate selectivity and self-inhibition in SLC4 anion transporters.
authors.affiliation: Molecular and Vascular Medicine Unit, Beth Israel Deaconess Med. Ctr., 330 Brookline Ave., Boston, MA 02215, USA.
authors.forename: Marina N
authors.initials: MN
authors.lastname: Chernova
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Stewart
authors.forename: Parul N
authors.initials: PN
authors.lastname: Barry
authors.forename: Michael L
authors.initials: ML
authors.lastname: Jennings
authors.forename: Seth L
authors.initials: SL
authors.lastname: Alper
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000838:Anions / Q000378:metabolism*; D001894:Borohydrides / Q000737:chemistry; D002712:Chlorides / Q000378:metabolism; D005260:Female; D018698:Glutamic Acid / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007555:Isoxazoles / Q000737:chemistry; D007700:Kinetics; D008698:Mesylates / Q000378:metabolism; D051379:Mice; D009865:Oocytes / Q000378:metabolism; D013431:Sulfates / Q000378:metabolism*; D013451:Sulfonic Acids / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D000838:Anions; D001894:Borohydrides; D002712:Chlorides; D007555:Isoxazoles; D008698:Mesylates; D013431:Sulfates; D013451:Sulfonic Acids; C045880:methanesulfonic acid; C009686:N-ethyl-5-phenylisoxazolium-3'-sulfonate; D018698:Glutamic Acid; C005741:sulfamic acid