The GPA-dependent, spherostomatocytosis mutant AE1 E758K induces GPA-independent, endogenous cation transport in amphibian oocytes

Andrew K. Stewart, David H. Vandorpe, John F. Heneghan, Fouad Chebib, Kathleen Stolpe, Arash Akhavein, E. Jennifer Edelman, Yelena Maksimova, Patrick G. Gallagher, Seth L. Alper

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Abstract

The previously undescribed heterozygous missense mutation E758K was discovered in the human AE1/SLC4A1/band 3 gene in two unrelated patients with well-compensated hereditary spherostomatocytic anemia (HSt). Oocyte surface expression of AE1 E758K, in contrast to that of wild-type AE1, required coexpressed glycophorin A (GPA). The mutant polypeptide exhibited, in parallel, strong GPA dependence of DIDS-sensitive 36Cl- influx, trans-anion-dependent 36Cl- efflux, and Cl -/HCO3- exchange activities at near wild-type levels. AE1 E758K expression was also associated with GPA-dependent increases of DIDS-sensitive pH-independent SO42- uptake and oxalate uptake with altered pH dependence. In marked contrast, the bumetanide- and ouabain-insensitive 86Rb+ influx associated with AE1 E758K expression was largely GPA-independent in Xenopus oocytes and completely GPA-independent in Ambystoma oocytes. AE1 E758K-associated currents in Xenopus oocytes also exhibited little or no GPA dependence. 86Rb+ influx was higher but inward cation current was lower in oocytes expressing AE1 E758K than previously reported in oocytes expressing the AE1 HSt mutants S731P and H734R. The pharmacological inhibition profile of AE1 E758K-associated 36Cl- influx differed from that of AE1 E758Kassociated 86Rb+ influx, as well as from that of wild-type AE1-mediated Cl- transport. Thus AE1 E758K-expressing oocytes displayed GPA-dependent surface polypeptide expression and anion transport, accompanied by substantially GPA-independent, pharmacologically distinct Rb + flux and by small, GPA-independent currents. The data strongly suggest that most of the increased cation transport associated with the novel HSt mutant AE1 E758K reflects activation of endogenous oocyte cation permeability pathways, rather than cation translocation through the mutant polypeptide.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume298
Issue number2
DOIs
StatePublished - Feb 1 2010
Externally publishedYes

Fingerprint

Glycophorin
Amphibians
Oocytes
Cations
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Anemia
Xenopus
Peptides
Anions
Ambystoma
Bumetanide
Oxalates
Ouabain
Missense Mutation
Permeability
Pharmacology

Keywords

  • Ambystoma oocytes
  • Chloride-bicarbonate exchange
  • Erythrocyte band 3
  • Glycophorin A
  • Xenopus oocytes

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

Cite this

The GPA-dependent, spherostomatocytosis mutant AE1 E758K induces GPA-independent, endogenous cation transport in amphibian oocytes. / Stewart, Andrew K.; Vandorpe, David H.; Heneghan, John F.; Chebib, Fouad; Stolpe, Kathleen; Akhavein, Arash; Edelman, E. Jennifer; Maksimova, Yelena; Gallagher, Patrick G.; Alper, Seth L.

In: American Journal of Physiology - Cell Physiology, Vol. 298, No. 2, 01.02.2010.

Research output: Contribution to journalArticle

Stewart, Andrew K. ; Vandorpe, David H. ; Heneghan, John F. ; Chebib, Fouad ; Stolpe, Kathleen ; Akhavein, Arash ; Edelman, E. Jennifer ; Maksimova, Yelena ; Gallagher, Patrick G. ; Alper, Seth L. / The GPA-dependent, spherostomatocytosis mutant AE1 E758K induces GPA-independent, endogenous cation transport in amphibian oocytes. In: American Journal of Physiology - Cell Physiology. 2010 ; Vol. 298, No. 2.
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abstract = "The previously undescribed heterozygous missense mutation E758K was discovered in the human AE1/SLC4A1/band 3 gene in two unrelated patients with well-compensated hereditary spherostomatocytic anemia (HSt). Oocyte surface expression of AE1 E758K, in contrast to that of wild-type AE1, required coexpressed glycophorin A (GPA). The mutant polypeptide exhibited, in parallel, strong GPA dependence of DIDS-sensitive 36Cl- influx, trans-anion-dependent 36Cl- efflux, and Cl -/HCO3- exchange activities at near wild-type levels. AE1 E758K expression was also associated with GPA-dependent increases of DIDS-sensitive pH-independent SO42- uptake and oxalate uptake with altered pH dependence. In marked contrast, the bumetanide- and ouabain-insensitive 86Rb+ influx associated with AE1 E758K expression was largely GPA-independent in Xenopus oocytes and completely GPA-independent in Ambystoma oocytes. AE1 E758K-associated currents in Xenopus oocytes also exhibited little or no GPA dependence. 86Rb+ influx was higher but inward cation current was lower in oocytes expressing AE1 E758K than previously reported in oocytes expressing the AE1 HSt mutants S731P and H734R. The pharmacological inhibition profile of AE1 E758K-associated 36Cl- influx differed from that of AE1 E758Kassociated 86Rb+ influx, as well as from that of wild-type AE1-mediated Cl- transport. Thus AE1 E758K-expressing oocytes displayed GPA-dependent surface polypeptide expression and anion transport, accompanied by substantially GPA-independent, pharmacologically distinct Rb + flux and by small, GPA-independent currents. The data strongly suggest that most of the increased cation transport associated with the novel HSt mutant AE1 E758K reflects activation of endogenous oocyte cation permeability pathways, rather than cation translocation through the mutant polypeptide.",
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T1 - The GPA-dependent, spherostomatocytosis mutant AE1 E758K induces GPA-independent, endogenous cation transport in amphibian oocytes

AU - Stewart, Andrew K.

AU - Vandorpe, David H.

AU - Heneghan, John F.

AU - Chebib, Fouad

AU - Stolpe, Kathleen

AU - Akhavein, Arash

AU - Edelman, E. Jennifer

AU - Maksimova, Yelena

AU - Gallagher, Patrick G.

AU - Alper, Seth L.

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N2 - The previously undescribed heterozygous missense mutation E758K was discovered in the human AE1/SLC4A1/band 3 gene in two unrelated patients with well-compensated hereditary spherostomatocytic anemia (HSt). Oocyte surface expression of AE1 E758K, in contrast to that of wild-type AE1, required coexpressed glycophorin A (GPA). The mutant polypeptide exhibited, in parallel, strong GPA dependence of DIDS-sensitive 36Cl- influx, trans-anion-dependent 36Cl- efflux, and Cl -/HCO3- exchange activities at near wild-type levels. AE1 E758K expression was also associated with GPA-dependent increases of DIDS-sensitive pH-independent SO42- uptake and oxalate uptake with altered pH dependence. In marked contrast, the bumetanide- and ouabain-insensitive 86Rb+ influx associated with AE1 E758K expression was largely GPA-independent in Xenopus oocytes and completely GPA-independent in Ambystoma oocytes. AE1 E758K-associated currents in Xenopus oocytes also exhibited little or no GPA dependence. 86Rb+ influx was higher but inward cation current was lower in oocytes expressing AE1 E758K than previously reported in oocytes expressing the AE1 HSt mutants S731P and H734R. The pharmacological inhibition profile of AE1 E758K-associated 36Cl- influx differed from that of AE1 E758Kassociated 86Rb+ influx, as well as from that of wild-type AE1-mediated Cl- transport. Thus AE1 E758K-expressing oocytes displayed GPA-dependent surface polypeptide expression and anion transport, accompanied by substantially GPA-independent, pharmacologically distinct Rb + flux and by small, GPA-independent currents. The data strongly suggest that most of the increased cation transport associated with the novel HSt mutant AE1 E758K reflects activation of endogenous oocyte cation permeability pathways, rather than cation translocation through the mutant polypeptide.

AB - The previously undescribed heterozygous missense mutation E758K was discovered in the human AE1/SLC4A1/band 3 gene in two unrelated patients with well-compensated hereditary spherostomatocytic anemia (HSt). Oocyte surface expression of AE1 E758K, in contrast to that of wild-type AE1, required coexpressed glycophorin A (GPA). The mutant polypeptide exhibited, in parallel, strong GPA dependence of DIDS-sensitive 36Cl- influx, trans-anion-dependent 36Cl- efflux, and Cl -/HCO3- exchange activities at near wild-type levels. AE1 E758K expression was also associated with GPA-dependent increases of DIDS-sensitive pH-independent SO42- uptake and oxalate uptake with altered pH dependence. In marked contrast, the bumetanide- and ouabain-insensitive 86Rb+ influx associated with AE1 E758K expression was largely GPA-independent in Xenopus oocytes and completely GPA-independent in Ambystoma oocytes. AE1 E758K-associated currents in Xenopus oocytes also exhibited little or no GPA dependence. 86Rb+ influx was higher but inward cation current was lower in oocytes expressing AE1 E758K than previously reported in oocytes expressing the AE1 HSt mutants S731P and H734R. The pharmacological inhibition profile of AE1 E758K-associated 36Cl- influx differed from that of AE1 E758Kassociated 86Rb+ influx, as well as from that of wild-type AE1-mediated Cl- transport. Thus AE1 E758K-expressing oocytes displayed GPA-dependent surface polypeptide expression and anion transport, accompanied by substantially GPA-independent, pharmacologically distinct Rb + flux and by small, GPA-independent currents. The data strongly suggest that most of the increased cation transport associated with the novel HSt mutant AE1 E758K reflects activation of endogenous oocyte cation permeability pathways, rather than cation translocation through the mutant polypeptide.

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KW - Chloride-bicarbonate exchange

KW - Erythrocyte band 3

KW - Glycophorin A

KW - Xenopus oocytes

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