Summary
Ouabain-resistant (OR), volume-or N-ethylmaleimide (NEM)-stimulated K+(Rb+)Cl− fluxes were measured in low-K+ sheep red cells and found to be functionally separate but immunologically similar. In anisosmotic solutions both K+ effluxes and Rb+ influxes of NEM-treated and control cells were additive. In contrast to the NEM-stimulated K+Cl− flux, metabolic depletion did not reduce K+Cl− flux of normal or swollen cells. The anion preference of OR K+ efflux in NEM-treated cells was Br−>Cl−>HCO −3 =F−≫I−=NO −3 =CNS−, and hence consistent with a reported Br−>Cl−>NO −3 sequence of the volume-dependent K+Cl− transport. Alloimmune anti-Ll antibodies known to decrease passive K+ fluxes in low K+ cells reduced by 51% both volume-and NEM-stimulated, furosemidesensitive Rb+Cl− fluxes suggesting their immunologic identity, a conclusion also supported by anti-L1 absorption studies. Since pretreatment with anti-L1 prevented the activation of Rb+ influx by NEM, and the impermeant glutathionmaleimide-I did not stimulate Rb+Cl− influx, the NEM reactive SH groups must be located apart from the L1 antigen either within the membrane or on its cytoplasmic face. A model is proposed consisting of a K+Cl− transport path(s) regulated by a protein with two functional subunits or domains; a chemically (C s) and a volume (V s)-stimulated domain, both interfacing with the L1 surface antigen. Attachment of alloanti-L1 from the outside reduces K+Cl− transport stimulated throughC s by NEM orV s by cell swelling.
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References
Abbott, R.E., Schachter, D. 1976. Impermeant maleimides. Oriented probes of erythrocyte membrane proteins.J. Biol. Chem. 251:7176–7183
Bauer, J., Lauf, P.K. 1983. Thiol-dependent passive K/Cl transport in sheep red cells: III. Differential reactivity of membrane SH groups with N-ethylmaleimide and iodoacetamide.J. Membrane Biol. 73:257–261
Becker, B.F., Duhm, J. 1978. Evidence for anionic cation transport of lithium, sodium, and potassium across the human erythrocyte membrane induced by divalent anions.J. Physiol. (London) 282:149–168
Brown, A.M., Ellory, J.C., Young, J.D., Lew, V.L. 1978. A calcium-activated potassium channel present in foetal red cells of the sheep but absent from reticulocytes and mature red cells.Biochim. Biophys. Acta 511:163–175
Cass, A., Dalmark, M. 1973. Equilibrium dialysis of ions in nystatin-treated red cells.Nature, New Biol. 244:47–49
Dani, J.A., Sanchez, J.A., Hille, B. 1983. Lyotropic anions. Na channel gating and Ca electrode response.J. Gen. Physiol. 82:255–281
Dunham, P.B. 1976. Anti-L serum. Two populations of antibodies affecting cation transport in LK erythrocytes of sheep and goats.Biochim. Biophys. Acta 443:219–226
Dunham, P.B. 1976. Passive potassium transport in LK sheep red cells. Effects of anti-L antibody and intracellular potassium.J. Gen. Physiol. 68:567–581
Dunham, P.B., Ellory, J.C. 1981. Passive potassium transport in low potassium sheep red cells: Dependence upon cell volume and chloride.J. Physiol. (London) 318:511–530
Eckel, R.E. 1958. Potassium exchange in human erythrocytes.J. Comp. Cell. Physiol. 51:81–108
Ellory, J.C., Dunham, P.B. 1980. Volume-dependent passive potassium transport in LK sheep red cells.In: Membrane Transport in Erythrocytes. Alfred Benzon Symposium 14, U.V. Lassen, H.H. Ussing and J.O. Wieth, editors. pp. 409–423. Munksgaard, Copenhagen
Funder, J., Tosteson, D.C., Wieth, J.O. 1978. Effects of bicarbonate on lithium transport in human red cells.J. Gen. Physiol. 71:721–746
Joiner, C.H., Lauf, P.K. 1978. Modulation of ouabain binding and potassium pump fluxes by cellular sodium and potassium in human and sheep erythrocytes.J. Physiol. (London) 283:177–196
Knauf, P.A., Ship, S., Breuer, W., McCulloch, L., Rothstein, A. 1978. Asymmetry of the red cell anion exchange system. Different mechanisms of reversible inhibition by N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate (NAP-Taurine) at the inside and outside of the membrane.J. Gen. Physiol. 72:607–630
Lauf, P.K. 1982. Active and passive cation transport and its association with membrane antigens in sheep erythrocytes. Developments and trends.In: Membranes and Transport. I. A.N. Martonosi, editor. pp. 553–558. Plenum, New York
Lauf, P.K. 1982. Kinetics of the SH-group dependent, chloride activated passive K+ transport in low K+ sheep red cells.Fed. Proc. 41:874
Lauf, P.K. 1983. Thiol-dependent passive K/Cl transport in sheep red cells: I. Dependence on chloride and external K+[Rb+] ions.J. Membrane Biol. 73:237–246
Lauf, P.K. 1983. Thiol-dependent passive K/Cl transport in sheep red cells: II. Loss of Cl− and N-ethylmaleimide sensitivity in maturing high K+ cells.J. Membrane Biol. 73:247–256.
Lauf, P.K. 1983. Thiol dependent, passive K/Cl transport in low K+ sheep red cells. IV. Furosemide inhibition as a function of external Rb+, Na+, and Cl−.J. Membrane Biol. 77:57–62
Lauf, P.K. 1983. Thiol-dependent passive K/Cl transport in sheep red blood cells. V. Dependence on metabolism.Am. J. Physiol. 245:C445-C448
Lauf, P.K. 1984. Immunological identity of K/Cl cotransport in low K+ sheep red cells stimulated by cell swelling or Nethylmaleimide.Biophys. J. 45:19a
Lauf, P.K., Adragna, N.C., Garay, R.P. 1984. Activation by N-ethylmaleimide of a latent K+−Cl− flux in human erythro cytes.Am. J. Physiol. 246:C385-C390
Lauf, P.K., Stiehl, B.J., Joiner, C.H. 1977. Active and passive cation transport and L antigen heterogeneity in low potassium sheep red cells. Evidence against the concept of leak-pump interconversion.J. Gen. Physiol. 70:221–242
Lauf, P.K., Theg, B.E. 1980. A chloride dependent K+ flux induced by N-ethylmaleimide in genetically low K+ sheep and goat erythrocytes.Biochem. Biophys. Res. Commun. 92:1422–1428
Lauf, P.K., Theg, B.E. 1981. N-ethylmaleimide enhances selectively passive K+ permeability in low potassium sheep red cells.In: Adv. Physiol. Sci. Vol. 6. Genetics, Structure and Function of Blood Cells. S.R. Hollan, G. Gardos and B. Sarkadi, editors. pp. 285–291. Pergamon, New York
Lauf, P.K., Zeidler, R.B., Kim, H.D. 1984. Pig reticulocytes: V. Development of Rb+ influx duringin vitro maturation.J. Cell. Physiol. (in press)
Logue, P., Anderson, C., Kanik, C., Farquharson, B., Dunham, P.B. 1983. Passive potassium transport in LK sheep red cells. Modification by N-ethylmaleimide.J. Gen. Physiol. 81:861–885
Szasz, I., Gardos, G. 1974. Mechanism of various drug effects on the Ca2+-dependent K+ efflux from human red blood cellsFEBS Lett. 44:213–216
Tolbert, A.B., Macey, R.I. 1982. The release of membranebound calcium by radiation and sulfhydryl reagents.J. Cell. Physiol. 79:43–52
Wiater, L.A., Dunham, P.B. 1983. Passive transport of K+ and Na+ in human red blood cells: Sulfhydryl binding agents and furosemide.Am. J. Physiol. 245:C348-C356
Wieth, J.O., Bjerrum, P.J. 1983. Transport and modifier sites in capnophorin, the anion transport protein of the erythrocyte membrane.In: Structure and Function of Membrane Proteins. E. Quagliariello and F. Palmieri, editors. pp. 95–106. Elsevier, Amsterdam
Wilbrandt, W. 1940. Die Abhángigkeit der Ionenpermeabilitát der Erythrocyten vom glykolytischen Stoffwechsel.Pfluegers Arch. Gesamte Physiol. 243:519–536
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Lauf, P.K. Thiol-dependent passive K+Cl− transport in sheep red blood cells: VI. Functional heterogeneity and immunologic identity with volume-stimulated K+(Rb+) fluxes. J. Membrain Biol. 82, 167–178 (1984). https://doi.org/10.1007/BF01868941
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DOI: https://doi.org/10.1007/BF01868941