Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 311K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JGP Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Chernova, M.N. Articles by Alper, S.L. Search for Related Content PubMed PubMed Citation Articles by Chernova, M.N. Articles by Alper, S.L. Social Bookmarking                            What's this?

Article

M.N. Chernova^*,, L. Jiang^*,, M. Crest^¶, M. Hand, D.H. Vandorpe^*, K. Strange, and S.L. Alper^*,,||||

From the ^* Molecular Medicine and Renal Units, Beth Israel Hospital; Critical Care Research Laboratories, Departments of Medicine (Nephrology) and Anesthesia, The Children's Hospital; Department of Medicine and ^|||| Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02215; and ^¶ Laboratoire de Neurobiologie, Centre National de la Recherche Scientifique, Marseille, France

Functional evaluation of chemically modified human erythrocytes has led to the proposal that amino acid residue E681 of the band 3 anion exchanger AE1 lies on the anion translocation pathway and is a proton carrier required for H⁺/SO₄^2– cotransport. We have tested in Xenopus oocytes the functional consequences of mutations in the corresponding residue E699 of mouse AE1. Most mutations tested abolished AE1-mediated Cl^– influx and efflux. Only the E699Q mutation increased stilbene disulfonate-sensitive efflux and influx of SO₄^2–. E699Q-mediated Cl^– influx was activated by elevation of intracellular SO₄^2–, but E699Q-mediated Cl^– efflux was undetectable. The DNDS (4,4'-dinitrostilbene-2,2'-disulfonic acid) sensitivity of E699Q-mediated SO₄^2– efflux was indistinguishable from that of wt AE1-mediated Cl^– efflux. The extracellular anion selectivity of E699Q-mediated SO₄^2– efflux was similar to that of wt AE1-mediated Cl^– efflux. The stoichiometry of E699Q-mediated exchange of extracellular Cl^– with intracellular SO₄^2– was 1:1. Whereas SO₄^2– injection into oocytes expressing wt AE1 produced little change in membrane potential or resistance, injection of SO₄^2–, but not of Cl^– or gluconate, into oocytes expressing E699Q depolarized the membrane by 17 mV and decreased membrane resistance by 66%. Replacement of bath Cl^– with isethionate caused a 28-mV hyperpolarization in SO₄^2–-loaded oocytes expressing E699Q, but had no effect on oocytes expressing wt AE1. Extracellular Cl^–-dependent depolarization of SO₄^2–-preloaded oocytes was blocked by DNDS. AE1 E699Q-mediated inward current measured in the presence of extracellular Cl^– was of magnitude sufficient to account for measured ³⁵SO₄^2– efflux. Thus, AE1 E699Q-mediated SO₄^2–/ Cl^– exchange operated largely, if not exclusively, as an electrogenic, asymmetric, 1:1 anion exchange. The data confirm the proposal that E699 resides on or contributes to the integrity of the anion translocation pathway of AE1. A single amino acid change in the sequence of AE1 converted electroneutral to electrogenic anion exchange without alteration of SO₄^2–/Cl^– exchange stoichiometry.

Key Words: band 3 • chloride/bicarbonate exchange • Xenopus oocyte • BCECF • sulfate

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				P. Dai, A. K. Stewart, F. Chebib, A. Hsu, J. Rozenfeld, D. Huang, D. Kang, V. Lip, H. Fang, H. Shao, et al. Distinct and novel SLC26A4/Pendrin mutations in Chinese and U.S. patients with nonsyndromic hearing loss Physiol Genomics, August 7, 2009; 38(3): 281 - 290. [Abstract] [Full Text] [PDF]

				S. L. Alper Molecular physiology and genetics of Na+-independent SLC4 anion exchangers J. Exp. Biol., June 1, 2009; 212(11): 1672 - 1683. [Abstract] [Full Text] [PDF]

				M. N. Chernova, A. K. Stewart, P. N. Barry, M. L. Jennings, and S. L. Alper Mouse Ae1 E699Q mediates SO42-i/aniono exchange with [SO42-]i-dependent reversal of wild-type pHo sensitivity Am J Physiol Cell Physiol, August 1, 2008; 295(2): C302 - C312. [Abstract] [Full Text] [PDF]

				A. K. Stewart, C. E. Kurschat, R. D. Vaughan-Jones, B. E. Shmukler, and S. L. Alper Acute regulation of mouse AE2 anion exchanger requires isoform-specific amino acid residues from most of the transmembrane domain J. Physiol., October 1, 2007; 584(1): 59 - 73. [Abstract] [Full Text] [PDF]

				A. Pushkin and I. Kurtz SLC4 base (HCO3-, CO32-) transporters: classification, function, structure, genetic diseases, and knockout models Am J Physiol Renal Physiol, March 1, 2006; 290(3): F580 - F599. [Abstract] [Full Text] [PDF]

				S. L Alper Molecular physiology of SLC4 anion exchangers Exp Physiol, January 1, 2006; 91(1): 153 - 161. [Abstract] [Full Text] [PDF]

				M. N. Chernova, D. H. Vandorpe, J. S. Clark, J. I. Williams, M. A. Zasloff, L. Jiang, and S. L. Alper Apparent receptor-mediated activation of Ca2+-dependent conductive Cl- transport by shark-derived polyaminosterols Am J Physiol Regulatory Integrative Comp Physiol, December 1, 2005; 289(6): R1644 - R1658. [Abstract] [Full Text] [PDF]

				B. E. Shmukler, C. E. Kurschat, G. E. Ackermann, L. Jiang, Y. Zhou, B. Barut, A. K. Stuart-Tilley, J. Zhao, L. I. Zon, I. A. Drummond, et al. Zebrafish slc4a2/ae2 anion exchanger: cDNA cloning, mapping, functional characterization, and localization Am J Physiol Renal Physiol, October 1, 2005; 289(4): F835 - F849. [Abstract] [Full Text] [PDF]

				T. Sherman, M. N. Chernova, J. S. Clark, L. Jiang, S. L. Alper, and K. Nehrke The abts and sulp families of anion transporters from Caenorhabditis elegans Am J Physiol Cell Physiol, August 1, 2005; 289(2): C341 - C351. [Abstract] [Full Text] [PDF]

				M. N. Chernova, L. Jiang, D. J. Friedman, R. B. Darman, H. Lohi, J. Kere, D. H. Vandorpe, and S. L. Alper Functional Comparison of Mouse slc26a6 Anion Exchanger with Human SLC26A6 Polypeptide Variants: DIFFERENCES IN ANION SELECTIVITY, REGULATION, AND ELECTROGENICITY J. Biol. Chem., March 4, 2005; 280(9): 8564 - 8580. [Abstract] [Full Text] [PDF]

				N. K. Dahl, L. Jiang, M. N. Chernova, A. K. Stuart-Tilley, B. E. Shmukler, and S. L. Alper Deficient HCO3- Transport in an AE1 Mutant with Normal Cl- Transport Can be Rescued by Carbonic Anhydrase II Presented on an Adjacent AE1 Protomer J. Biol. Chem., November 7, 2003; 278(45): 44949 - 44958. [Abstract] [Full Text] [PDF]

				M. N Chernova, L. Jiang, B. E Shmukler, C. W Schweinfest, P. Blanco, S. D Freedman, A. K Stewart, and S. L Alper Acute regulation of the SLC26A3 congenital chloride diarrhoea anion exchanger (DRA) expressed in Xenopus oocytes J. Physiol., May 15, 2003; 549(1): 3 - 19. [Abstract] [Full Text] [PDF]

				M. N. Chernova, A. K. Stewart, L. Jiang, D. J. Friedman, Y. Z. Kunes, and S. L. Alper Structure-function relationships of AE2 regulation by Ca2+i-sensitive stimulators NH+4 and hypertonicity Am J Physiol Cell Physiol, May 1, 2003; 284(5): C1235 - C1246. [Abstract] [Full Text] [PDF]

				A.K. Stewart, M.N. Chernova, B.E. Shmukler, S. Wilhelm, and S.L. Alper Regulation of AE2-mediated Cl- Transport by Intracellular or by Extracellular pH Requires Highly Conserved Amino Acid Residues of the AE2 NH2-terminal Cytoplasmic Domain J. Gen. Physiol., October 29, 2002; 120(5): 707 - 722. [Abstract] [Full Text] [PDF]

				Q. Xie, R. Welch, A. Mercado, M. F. Romero, and D. B. Mount Molecular characterization of the murine Slc26a6 anion exchanger: functional comparison with Slc26a1 Am J Physiol Renal Physiol, October 1, 2002; 283(4): F826 - F838. [Abstract] [Full Text] [PDF]

				G. G. Goss, L. Jiang, D. H. Vandorpe, D. Kieller, M. N. Chernova, M. Robertson, and S. L. Alper Role of JNK in hypertonic activation of Cl--dependent Na+/H+ exchange in Xenopus oocytes Am J Physiol Cell Physiol, December 1, 2001; 281(6): C1978 - C1990. [Abstract] [Full Text] [PDF]

				S. Casula, B. E. Shmukler, S. Wilhelm, A. K. Stuart-Tilley, W. Su, M. N. Chernova, C. Brugnara, and S. L. Alper A Dominant Negative Mutant of the KCC1 K-Cl Cotransporter. BOTH N- AND C-TERMINAL CYTOPLASMIC DOMAINS ARE REQUIRED FOR K-Cl COTRANSPORT ACTIVITY J. Biol. Chem., November 2, 2001; 276(45): 41870 - 41878. [Abstract] [Full Text] [PDF]

				R. Zhao and R. A. F. Reithmeier Expression and characterization of the anion transporter homologue YNL275w in Saccharomyces cerevisiae Am J Physiol Cell Physiol, July 1, 2001; 281(1): C33 - C45. [Abstract] [Full Text] [PDF]

				R. Beckmann, J. S. Smythe, D. J. Anstee, and M. J. A. Tanner Coexpression of band 3 mutants and Rh polypeptides: differential effects of band 3 on the expression of the Rh complex containing D polypeptide and the Rh complex containing CcEe polypeptide Blood, April 15, 2001; 97(8): 2496 - 2505. [Abstract] [Full Text] [PDF]

				D. Stakisaitis, M. S. Lapointe, and D. Batlle Mechanisms of chloride transport in thymic lymphocytes Am J Physiol Renal Physiol, February 1, 2001; 280(2): F314 - F324. [Abstract] [Full Text] [PDF]

				P. A. Knauf, N. M. Raha, and L. J. Spinelli The Noncompetitive Inhibitor Ww781 Senses Changes in Erythrocyte Anion Exchanger (Ae1) Transport Site Conformation and Substrate Binding J. Gen. Physiol., February 1, 2000; 115(2): 159 - 174. [Abstract] [Full Text] [PDF]

				W. Su, B. E. Shmukler, M. N. Chernova, A. K. Stuart-Tilley, L. de Franceschi, C. Brugnara, and S. L. Alper Mouse K-Cl cotransporter KCC1: cloning, mapping, pathological expression, and functional regulation Am J Physiol Cell Physiol, November 1, 1999; 277(5): C899 - C912. [Abstract] [Full Text] [PDF]

				S. Bahar, C. T. Gunter, C. Wu, S. D. Kennedy, and P. A. Knauf Persistence of external chloride and DIDS binding after chemical modification of Glu-681 in human band 3 Am J Physiol Cell Physiol, October 1, 1999; 277(4): C791 - C799. [Abstract] [Full Text] [PDF]

				X.-B. Tang, M. Kovacs, D. Sterling, and J. R. Casey Identification of Residues Lining the Translocation Pore of Human AE1, Plasma Membrane Anion Exchange Protein J. Biol. Chem., February 5, 1999; 274(6): 3557 - 3564. [Abstract] [Full Text] [PDF]

				R. T. Timmer and R. B. Gunn Inducible expression of erythrocyte band 3 protein Am J Physiol Cell Physiol, January 1, 1999; 276(1): C66 - C75. [Abstract] [Full Text] [PDF]

				X.-B. Tang, J. Fujinaga, R. Kopito, and J. R. Casey Topology of the Region Surrounding Glu681 of Human AE1 Protein, the Erythrocyte Anion Exchanger J. Biol. Chem., August 28, 1998; 273(35): 22545 - 22553. [Abstract] [Full Text] [PDF]

				D. H. Vandorpe, B. E. Shmukler, L. Jiang, B. Lim, J. Maylie, J. P. Adelman, L. de Franceschi, M. D. Cappellini, C. Brugnara, and S. L. Alper cDNA Cloning and Functional Characterization of the Mouse Ca2+-gated K+ Channel, mIK1. ROLES IN REGULATORY VOLUME DECREASE AND ERYTHROID DIFFERENTIATION J. Biol. Chem., August 21, 1998; 273(34): 21542 - 21553. [Abstract] [Full Text] [PDF]

				P. Jarolim, C. Shayakul, D. Prabakaran, L. Jiang, A. Stuart-Tilley, H. L. Rubin, S. Simova, J. Zavadil, J. T. Herrin, J. Brouillette, et al. Autosomal Dominant Distal Renal Tubular Acidosis Is Associated in Three Families with Heterozygosity for the R589H Mutation in the AE1 (Band 3) Cl-/HCO3- Exchanger J. Biol. Chem., March 13, 1998; 273(11): 6380 - 6388. [Abstract] [Full Text] [PDF]

				D. H. Vandorpe, M. N. Chernova, L. Jiang, L. K. Sellin, S. Wilhelm, A. K. Stuart-Tilley, G. Walz, and S. L. Alper The Cytoplasmic C-terminal Fragment of Polycystin-1 Regulates a Ca2+-permeable Cation Channel J. Biol. Chem., February 2, 2001; 276(6): 4093 - 4101. [Abstract] [Full Text] [PDF]

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents