Abstract
Passive K transport, as modified by N-ethyl maleimide (NEM), was studied in erythrocytes of the low-K (LK) phenotype of sheep. Brief (5- min) treatment with NEM at less than 0.5 mM caused inhibition of passive K influx; NEM at concentrations greater than 0.5 mM caused stimulation of K influx. NEM had similar effects on K efflux. The treatments with NEM did not affect cell volumes (passive K transport in LK cells is sensitive to changes in cell volume). The stimulation of K transport by high [NEM] was also not a consequence of an effect on the metabolic state of the cells. Passive K transport in LK cells is dependent on Cl (it is inhibited in Cl-free media; it may be K/Cl cotransport). NEM had no effect on K influx in Cl-free (NO3- substituted) media. Pretreatment of the cells with anti-L antiserum (L antigen is found on LK cells and not on HK cells) prevented stimulation of K influx by NEM, but did not prevent inhibition. Therefore, NEM modifies the Cl-dependent K transport pathway at two separate sites, a low-affinity site, at which it stimulates, and a high-affinity site, at which it inhibits. Anti-L antibody prevents NEM's action, but only at the low-affinity site.
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- Aickin C. C., Deisz R. A., Lux H. D. Ammonium action on post-synaptic inhibition in crayfish neurones: implications for the mechanism of chloride extrusion. J Physiol. 1982 Aug;329:319–339. doi: 10.1113/jphysiol.1982.sp014305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BENESCH R., BENESCH R. E. Determination of--SH groups in proteins. Methods Biochem Anal. 1962;10:43–70. doi: 10.1002/9780470110270.ch2. [DOI] [PubMed] [Google Scholar]
- Beauge L. A., Adragna N. The kinetics of ouabain inhibition and the partition of rubidium influx in human red blood cells. J Gen Physiol. 1971 May;57(5):576–592. doi: 10.1085/jgp.57.5.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brooker R. J., Slayman C. W. [14C]N-ethylmaleimide labeling of the plasma membrane [H+]-ATPase of Neurospora crassa. J Biol Chem. 1983 Jan 10;258(1):222–226. [PubMed] [Google Scholar]
- Chipperfield A. R. Chloride dependence of frusemide- and phloretin-sensitive passive sodium and potassium fluxes in human red cells. J Physiol. 1981 Mar;312:435–444. doi: 10.1113/jphysiol.1981.sp013636. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Duhm J., Becker B. F. Studies on lithium transport across the red cell membrane. V. On the nature of the Na+-dependent Li+ countertransport system of mammalian erythrocytes. J Membr Biol. 1979 Dec 31;51(3-4):263–286. doi: 10.1007/BF01869087. [DOI] [PubMed] [Google Scholar]
- Dunham P. B. Anti-L serum. Two populations of antibodies affecting cation transport in LK erythrocytes of sheep and goats. Biochim Biophys Acta. 1976 Aug 16;443(2):219–226. doi: 10.1016/0005-2736(76)90505-8. [DOI] [PubMed] [Google Scholar]
- Dunham P. B., Ellory J. C. Passive potassium transport in low potassium sheep red cells: dependence upon cell volume and chloride. J Physiol. 1981 Sep;318:511–530. doi: 10.1113/jphysiol.1981.sp013881. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunham P. B., Ellory J. C. Stimulation of the sodium-potassium pump by trypsin in low potassium type erythrocytes of goats. J Physiol. 1980 Apr;301:25–37. doi: 10.1113/jphysiol.1980.sp013185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunham P. B., Hoffman J. F. Active cation transport and ouabain binding in high potassium and low potassium red blood cells of sheep. J Gen Physiol. 1971 Jul;58(1):94–116. doi: 10.1085/jgp.58.1.94. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunham P. B. Passive potassium transport in LK sheep red cells. Effects of anti-L antibody and intracellular potassium. J Gen Physiol. 1976 Dec;68(6):567–581. doi: 10.1085/jgp.68.6.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunham P. B., Stewart G. W., Ellory J. C. Chloride-activated passive potassium transport in human erythrocytes. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1711–1715. doi: 10.1073/pnas.77.3.1711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ellory J. C., Dunham P. B., Logue P. J., Stewart G. W. Anion-dependent cation transport in erythrocytes. Philos Trans R Soc Lond B Biol Sci. 1982 Dec 1;299(1097):483–495. doi: 10.1098/rstb.1982.0146. [DOI] [PubMed] [Google Scholar]
- Ellory J. C., Tucker E. M. Stimulation of the potassium transport system in low potassium type sheep red cells by a specific antigen antibody reaction. Nature. 1969 May 3;222(5192):477–478. doi: 10.1038/222477a0. [DOI] [PubMed] [Google Scholar]
- Ericson A. C., Spring K. R. Coupled NaCl entry into Necturus gallbladder epithelial cells. Am J Physiol. 1982 Sep;243(3):C140–C145. doi: 10.1152/ajpcell.1982.243.3.C140. [DOI] [PubMed] [Google Scholar]
- Ericson A. C., Spring K. R. Volume regulation by Necturus gallbladder: apical Na+-H+ and Cl(-)-HCO-3 exchange. Am J Physiol. 1982 Sep;243(3):C146–C150. doi: 10.1152/ajpcell.1982.243.3.C146. [DOI] [PubMed] [Google Scholar]
- GLYNN I. M. The action of cardiac glycosides on sodium and potassium movements in human red cells. J Physiol. 1957 Apr 3;136(1):148–173. doi: 10.1113/jphysiol.1957.sp005749. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garrahan P. J., Glynn I. M. Facftors affecting the relative magnitudes of the sodium:potassium and sodium:sodium exchanges catalysed by the sodium pump. J Physiol. 1967 Sep;192(1):189–216. doi: 10.1113/jphysiol.1967.sp008296. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geck P., Pietrzyk C., Burckhardt B. C., Pfeiffer B., Heinz E. Electrically silent cotransport on Na+, K+ and Cl- in Ehrlich cells. Biochim Biophys Acta. 1980 Aug 4;600(2):432–447. doi: 10.1016/0005-2736(80)90446-0. [DOI] [PubMed] [Google Scholar]
- Glynn I. M., Lew V. L., Lüthi U. Reversal of the potassium entry mechanism in red cells, with and without reversal of the entire pump cycle. J Physiol. 1970 Apr;207(2):371–391. doi: 10.1113/jphysiol.1970.sp009067. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greger R., Schlatter E. Presence of luminal K+, a prerequisite for active NaCl transport in the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflugers Arch. 1981 Nov;392(1):92–94. doi: 10.1007/BF00584588. [DOI] [PubMed] [Google Scholar]
- Haas M., Schmidt W. F., 3rd, McManus T. J. Catecholamine-stimulated ion transport in duck red cells. Gradient effects in electrically neutral [Na + K + 2Cl] Co-transport. J Gen Physiol. 1982 Jul;80(1):125–147. doi: 10.1085/jgp.80.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoffman J. F., Kregenow F. M. The characterization of new energy dependent cation transport processes in red blood cells. Ann N Y Acad Sci. 1966 Jul 14;137(2):566–576. doi: 10.1111/j.1749-6632.1966.tb50182.x. [DOI] [PubMed] [Google Scholar]
- Imler J. R., Vidaver G. A. Anion effects on glycine entry into pigeon red blood cells. Biochim Biophys Acta. 1972 Oct 23;288(1):153–165. doi: 10.1016/0005-2736(72)90233-7. [DOI] [PubMed] [Google Scholar]
- Kregenow F. M. Osmoregulatory salt transporting mechanisms: control of cell volume in anisotonic media. Annu Rev Physiol. 1981;43:493–505. doi: 10.1146/annurev.ph.43.030181.002425. [DOI] [PubMed] [Google Scholar]
- Lauf P. K., Theg B. E. A chloride dependent K+ flux induced by N-ethylmaleimide in genetically low K+ sheep and goat erythrocytes. Biochem Biophys Res Commun. 1980 Feb 27;92(4):1422–1428. doi: 10.1016/0006-291x(80)90445-3. [DOI] [PubMed] [Google Scholar]
- Lubowitz H., Whittam R. Ion movements in human red cells independent of the sodium pump. J Physiol. 1969 May;202(1):111–131. doi: 10.1113/jphysiol.1969.sp008798. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McRoberts J. A., Erlinger S., Rindler M. J., Saier M. H., Jr Furosemide-sensitive salt transport in the Madin-Darby canine kidney cell line. Evidence for the cotransport of Na+, K+, and Cl-. J Biol Chem. 1982 Mar 10;257(5):2260–2266. [PubMed] [Google Scholar]
- Musch M. W., Orellana S. A., Kimberg L. S., Field M., Halm D. R., Krasny E. J., Jr, Frizzell R. A. Na+-K+-Cl- co-transport in the intestine of a marine teleost. Nature. 1982 Nov 25;300(5890):351–353. doi: 10.1038/300351a0. [DOI] [PubMed] [Google Scholar]
- Rao A., Reithmeier R. A. Reactive sulfhydryl groups of the band 3 polypeptide from human erythroycte membranes. Location in the primary structure. J Biol Chem. 1979 Jul 10;254(13):6144–6150. [PubMed] [Google Scholar]
- Rasmusen B. A., Hall J. G. Association between potassium concentration and serological type of sheep red blood cells. Science. 1966 Mar 25;151(3717):1551–1552. doi: 10.1126/science.151.3717.1551. [DOI] [PubMed] [Google Scholar]
- Russell J. M. Chloride and sodium influx: a coupled uptake mechanism in the squid giant axon. J Gen Physiol. 1979 Jun;73(6):801–818. doi: 10.1085/jgp.73.6.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sachs J. R. Ouabain-insensitive sodium movements in the human red blood cell. J Gen Physiol. 1971 Mar;57(3):259–282. doi: 10.1085/jgp.57.3.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- TOSTESON D. C., HOFFMAN J. F. Regulation of cell volume by active cation transport in high and low potassium sheep red cells. J Gen Physiol. 1960 Sep;44:169–194. doi: 10.1085/jgp.44.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
- TUCKER E. M. FURTHER OBSERVATIONS ON THE I BLOOD GROUP IN SHEEP. Vox Sang. 1965 Mar-Apr;10:195–205. doi: 10.1111/j.1423-0410.1965.tb04337.x. [DOI] [PubMed] [Google Scholar]
- Wiley J. S., Cooper R. A. A furosemide-sensitive cotransport of sodium plus potassium in the human red cell. J Clin Invest. 1974 Mar;53(3):745–755. doi: 10.1172/JCI107613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winslow J. W. The reaction of sulfhydryl groups of sodium and potassium ion-activated adenosine triphosphatase with N-ethylmaleimide. The relationship between ligand-dependent alterations of nucleophilicity and enzymatic conformational states. J Biol Chem. 1981 Sep 25;256(18):9522–9531. [PubMed] [Google Scholar]