Skip to main content
Log in

Sodium-dependentl-serine transport in plasma membrane vesicles isolated from Ehrlich cells by two-phase compartmentation

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Plasma membrane vesicles were prepared from Ehrlich cells using two-phase system compartmentation. The highly pure plasma membrane vesicles obtained presented a negligible mitochondrial contamination and were suitable for studies of amino acid transport.l-Serine transport showed a clear ionic specificity, maximum incorporation being observed when an inwardly directed NaSCN gradient was used. Na+-dependentl-serine transport was dependent on assay temperature and membrane potential, and it seemed to be carried out by two different transport systems. An essential sulfhydryl group seemed to be involved in the transport process.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baker, N., Sandborg, C., Morris, D., Ookhtens, M. 1974. Competition for host essential and nonessential fatty acid in Ehrlich ascites carcinoma in mice.Cancer Res. 37:2218–2225

    Google Scholar 

  • Bardin, C., Johnstone, R.M. 1978. Sodium-dependent amino acid transport in reconstituted membrane vesicles from Ehrlich ascites cell plasma membranes.J. Biol. Chem. 253:1725–1732

    Google Scholar 

  • Biber, J., Haüser, H. 1979. The role of SH-group in the concentrative transport ofd-glucose into brush border membrane vesicles.FEBS Lett. 108:451–456

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.anal. Biochem. 72:248–254

    Google Scholar 

  • Christensen, H.N. 1989. Distinguishing amino acid transport system of a given cell or tissue.Methods Enzymol. 173:576–616

    Google Scholar 

  • Christensen, H.N. 1990. Role of amino acid transport and countertransport in nutrition and metabolism.Physiol. Rev. 70:43–77

    Google Scholar 

  • Christensen, H.N., Liang, M., Archerd, E.G. 1967. A distinct Na+-requiring transport system for alanine, serine, cysteine, and similar amino acids.J. Biol. Chem. 242:5237–5246

    Google Scholar 

  • Christensen, H.N., Oxender, D.L., Liang, M., Vatz, K.A. 1965. The use of N-methylation to direct the route of mediated transport of amino acid.J. Biol. Chem. 240:3609–3616

    Google Scholar 

  • Colombini, M., Johnstone, R.M. 1973. Preparation and properties of the (Na++K+)-ATPase of plasma membranes from Ehrlich ascites cells.Biochim. Biophys. Acta 323:69–86

    Google Scholar 

  • Colombini, M., Johnstone, R.M. 1974. Na+-gradient-stimulated AIB transport in membrane vesicles from Ehrlich ascites cells.J. Membrane Biol. 18:315–334

    Google Scholar 

  • Driessen, A.J.M., Konings, W.N. 1990. Reactive exofacial sulfhydryl-groups on the arginine-ornithine antiporter ofLactococcus lactis.Biochim. Biophys. Acta 1015:87–95

    Google Scholar 

  • Esmann, M. 1988. ATPase and phosphatase activity of Na+, K+-ATPase: Molar and specific activity, protein determination.Methods Enzymol. 156:105–115

    Google Scholar 

  • Flanagan, S.D. 1985. Partitioning of animal membranes and organelles.In: Partitioning in Aqueous Two-Phase Systems. H. Walter, D.E. Brooks, and D. Fisher, editors. pp. 453–495. Academic, London

    Google Scholar 

  • Gruber, M., Cheng, K., Lepock, J.R., Thompson, J.E. 1984. Improved yield of plasma membrane from mammalian cells through modifications of two-phase polymer isolation procedure.Anal. Biochem. 138:102–118

    Google Scholar 

  • Handlogten, M.E., García-Cañero, R., Lancaster, K.T., Christensen, H.N. 1981. Surprising differences in substrate selectivity and other properties of system A and ASC between rat hepatocytes and the hepatoma cell line HTC.J. Biol. Chem. 256:7905–7909

    Google Scholar 

  • Hanning, K., Heidrich, H.G. 1974. The use of continuous preparative free-flow electrophoresis for dissociating cell fractions and isolation of membranous components.Methods Enzymol. 31:746–761

    Google Scholar 

  • Herrero, E., Giménez, C., Aragón, M.C. 1987. Chronic administration of lithium modulates tryptophan transport by changing the properties of the synaptosomal plasma membrane.Life Sci. 41:643–650

    Google Scholar 

  • Holley, R.W. 1972. Unifying hypothesis concerning the nature of malignant growth.Proc. Natl. Acad. Sci. USA 69:2840–2841

    Google Scholar 

  • Im, W.B., Spector, A.A. 1980. Sodium-dependent neutral amino acid transport in native and reconstituted membrane vesicles from Ehrlich cells.J. Biol. Chem. 255:764–770

    Google Scholar 

  • Kashiwamata, S., Goto, S., Semba, R., Suzuki, F. 1979. Inhibition by bilirubin of (sodium-potassium ion)-activated adenosine triphosphatase and potassium ion-activatedp-nitrophenylphosphatase activities of sodium iodide-treated microsomes from young rat cerebrum.J. Biol. Chem. 254:4577–4584

    Google Scholar 

  • Kilberg, M.S., Christensen, H.N. 1979. Electron-transferring enzymes in the plasma membrane of the Ehrlich ascites tumor cell.Biochemistry 18:1525–1530

    Google Scholar 

  • Knox, W.E., Herzfeld, A., Hodson, J. 1969. Phosphoserine phosphatase distribution in normal and neoplastic rat tissue.Arch. Biochem. Biophys. 132:397–405

    Google Scholar 

  • Krause, R., James, J.H., Humphrey, C., Fisher, J.E. 1979. Plasma and brain amino acids in Walker 256 carcinosarcoma-bearing rats.Cancer Res. 39:3065–3069

    Google Scholar 

  • Larsson, C. 1983. Partition in aqueous two-phase systems.In: Isolation of Membranes and Organelles from Plant Cells. J.L. Hall and A.L. Moore, editors. pp. 277–309. Academic, London

    Google Scholar 

  • Larsson, C., Andersson, B. 1979. Two-phase methods for chloroplasts, chloroplast elements and mitochondria.In: Plant Organelles: Methodological Surveys. B. Biochemistry, E. Reid, editor. Vol. 9, pp. 498–527. Academic, London

    Google Scholar 

  • Lazo, P. 1981. Amino acid and glucose utilization by different metabolic pathways in ascites tumour cells.Eur. J. Biochem. 117:19–25

    Google Scholar 

  • Levin, L., Gevers, W. 1981a. Metabolic alterations in cancer: I.S.A. Med. J. 59:518–521

    Google Scholar 

  • Levin, L., Gevers, W. 1981b. Metabolic alterations in cancer: II.S.A. Med. J. 59:553–556

    Google Scholar 

  • Lücke, H., Haase, W., Murer, H. 1977. Amino acid transport in brush-border-membrane vesicles isolated from human small intestine.Biochem. J. 168:529–532

    Google Scholar 

  • Lynch, A.M., McGivan, J.D. 1987. Evidence for a single common sodium-dependent transport system for alanine, glutamine, leucine, and phenylalanine in brush-border membrane vesicles from bovine kidney.Biochim. Biophys. Acta 899:176–184

    Google Scholar 

  • Makowske, M., Christensen, H.N. 1982. Hepatic transport system interconverted by protonation from service for neutral to service for anionic amino acids.J. Biol. Chem. 257:14635–14638

    Google Scholar 

  • Márquez, J., Quesada, A.R., Sánchez-Jiménez, F., Núñez de Castro, I. 1986. Determination of 27 dansyl-amino acid derivatives in biological fluids by reversed-phase high-performance liquid chromatography.J. Chromatogr. Biomed. Appl. 380:275–283

    Google Scholar 

  • Márquez, J., Sánchez-Jiménez, F., Medina, M.A., Quesada, A.R., Núñez de Castro, I. 1989. Nitrogen metabolism in tumor bearing mice.Arch. Biochem. Biophys. 268:667–675

    Google Scholar 

  • McCormick, J., Johnstone, R.M. 1990. Evidence for an essential sulfhydryl group at the substrate binding site of the A system transport of Ehrlich cell plasma membranes.Biochem. Cell Biol. 68:512–519

    Google Scholar 

  • McCormick, J., Tsang, D., Johnstone, R.M. 1984. A simple and efficient method for reconstitution of amino acid and glucose transport systems from Ehrlich ascites cells.Arch. Biochem. Biophys. 231:355–365

    Google Scholar 

  • Medina, M.A. 1989. Glutamina y Tumor: Transporte y Metabolismo. Ph.D. Thesis, Málaga University (in Spanish)

  • Medina, M.A., Núñez de Castro, I. 1990a. Glutaminolysis and glycolysis interactions in proliferant cells.Int. J. Biochem. 22:681–683

    Google Scholar 

  • Medina, M.A., Núñez de Castro, I. 1990b. Plasma membrane oxidoreduction and amino acid transport.In: Oxidoreduction in the Plasma Membrane: Relation to Growth and Transport. Vol. 1. pp. 247–255. F.L. Crane, D.J. Morré, and H. Löw, editors. CRC, Boca Ratón

    Google Scholar 

  • Nordlie, R.C., Arion, W.J. 1966. Glucose-6-phosphatase.Methods Enzymol. 9:619–625

    Google Scholar 

  • Ookhtens, M., Baker, N. 1979. Fatty acid oxidation to H2O by Ehrlich ascites carcinoma in mice.Cancer Res. 39:973–980

    Google Scholar 

  • Perán, S., Muñoz, M., Sainz, M.T. 1990a. Rapid and steady-state amino acid transport in perfused human fibroblast and colon adenocarcinoma cells: Effects of methotrexate.Biochim. Biophys. Acta 1024:233–240

    Google Scholar 

  • Perán, S., Muñoz, M., Sainz, M.T. 1990b. New approach to the study of transport of biomolecules by microcarrier cell cultures perfused in column applying a high resolution paired tracer technique.In: Cell Membrane Transport: Experimental Approaches and Methodologies. D.L. Yudilevich, I. Cabanchik, R. Devés, and S. Perán, editors. Plenum, London (in press)

    Google Scholar 

  • Pola, E., Beltrán, J., Roca, A., Palacín, M., Zorzano, A., Testar, X. 1990. Sensitivity of system A and ASC transport activities to thiol-group-modifying reagents in rat liver plasma-membrane vesicles.Biochem. J. 271:297–303

    Google Scholar 

  • Quesada, A.R., McGivan, J.D. 1988. A rapid method for the functional reconstitution of amino acid transport system from rat liver plasma membranes.Biochem. J. 255:963–969

    Google Scholar 

  • Quesada, A.R., Medina, M.A., Márquez, J., Sánchez-Jiménez, F., Núñez de Castro, I. 1988. Contribution by host tissues to circulating glutamine in mice inoculated with Ehrlich ascites tumor cells.Cancer Res. 48:1551–1553

    Google Scholar 

  • Sauer, L.A., Dauchy, R.T. 1983. Ketone body, glucose, lactic acid and amino acid utilization by tumorsin vivo in fasted rats.Cancer Res. 43:3497–3503

    Google Scholar 

  • Sips, H.J., De Graaf, P.A., Van Dam, K. 1982. Transport ofl-aspartate andl-glutamate in plasma-membrane vesicles from rat liver.Eur. J. Biochem. 122:259–264

    Google Scholar 

  • Sips, H.J., Van Amelsvoort, J.M.M., Van Dam, K. 1980. Amino acid transport in plasma membrane vesicles from rat liver.Eur. J. Biochem. 105:217–224

    Google Scholar 

  • Snell, K. 1985. Enzymes of serine metabolism in normal and neoplastic tissues.Biochem. Biophys. Acta 843:276–281

    Google Scholar 

  • Spector, A.A., Steinberg, D. 1965. The utilization of unsterified palmitate by Ehrlich ascites tumor cells.J. Biol. Chem. 240:3747–3753

    Google Scholar 

  • Thordinke, J., Pelliniemi, T.T., Beck, W.S. 1979. Serine hydroxymethyltransferase activity and serine incorporation in leukocytes.Cancer Res. 39:3435–3440

    Google Scholar 

  • Urdiales, J.L., Medina, M.A., Núñez de Castro, I., Sánchez-Jiménez, F. 1989. Early systemic effect on the hepatic mitochondria of tumour bearing mice.Cancer Lett. 44:179–183

    Google Scholar 

  • Van Amelsvoort, J.M.M., Sips, H.J., Van Dam, K. 1978. Sodium-dependent alanine transport in plasma membrane vesicles from rat liver.Biochem. J. 174:1083–1086

    Google Scholar 

  • Zafra, F., Giménez, C. 1986. Characterization of glycine uptake in plasma membrane vesicles isolated from cultured glioblastoma cells.Brain Res. 397:108–116

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Luque, P., Márquez, J., Núñez de Castro, I. et al. Sodium-dependentl-serine transport in plasma membrane vesicles isolated from Ehrlich cells by two-phase compartmentation. J. Membrain Biol. 123, 247–254 (1991). https://doi.org/10.1007/BF01870407

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01870407

Key Words

Navigation