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Recognition of sodium- and potassium-dependent adenosine triphosphatase on mouse lymphoid cells by means of a monoclonal antibody

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Summary

Previous evidence has established the similarity between (Na++K+)-ATPase (ATP phosphohydrolase, EC.3.6.1.3) and the antigen recognized by the rat antimouse monoclonal antibody anti-BSP-3. This antibody has been used for investigation of the surface expression and biochemical analysis of the enzyme in different mouse lymphoid populations. The BSP-3 determinant is found on almost all thymocytes and concanavalin A-induced thymocytes, to a lesser extent on bone marrow cells and also on a minor population of spleen cells. Spleen cells from athymic mice are negative. The (Na++ K+)-ATPase purified from mouse thymus by affinity chromatography migrates in SDS-polyacrylamide gels in the form of two polypeptide chains of 105000 and 51000 daltons. Chains of the same molecular weight, fractionated on SDS-PAGE from microsomes of mouse thymuses, are shown to react with subunit-specific polyclonal antisera against ATPase in immunoblotting experiments. Immunoprecipitation with anti-BSP-3 from surface iodinated thymocytes yields only the small subunit. Comparison of the chains isolated from thymus and brain shows molecular weight differences in both subunits. These results, and variations in the reactivity pattern of the anti-BSP-3 antibody on several cell types, may indicate a possible heterogeneity of the (Na++K+)ATPase expressed by various tissues and cells.

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References

  • Averdunk R, Müller J, Wenzel B (1976) Untersuchungen über den Mechanismus der Aktivierung der Lymphocytenmembran-ATPasen durch Concanavalin A. J Clin Chem Clin Biochem 14:339–344

    Google Scholar 

  • Ball WJ, Schwartz A, Lessard JL (1982) Isolation and characterzation of monoclonal antibodies to (Na+ +K+)-ATPase. Biochim Biophys Acta 719:413–423

    Google Scholar 

  • Ball WJ Jr, Collins JH, Lane LK, Schwartz A (1983) Studies on the antigenic properties of the catalytic and glycoprotein subunits of Na+, K+-ATPase. Arch Biochem Biophys 221:371–380

    Google Scholar 

  • Barber BH, Delovitch TH (1979) The identification of actin as a major lymphocyte component. J Immunol 122:320–325

    Google Scholar 

  • Bridgen J, Snary D, Crumpton MJ, Barnstable CJ, Goodfellow PN, Bodmer WF (1976) Isolation and N-terminal amino acid sequence of membrane-bound human HLA-A and HLA-B antigens. Nature (Lond) 261:200–205

    Google Scholar 

  • Brown WRA, Barclay AN, Sunderland CA, Williams AF (1981) Identification of a glycoprotein-like molecule at the cell surface of rat thymocytes. Nature (Lond) 289:456–460

    Google Scholar 

  • Bruce J, Symington FW, McKearn TJ, Sprent J (1981) A monoclonal antibody discriminating between subsets of T and B cells. J Immunol 127:2496–2501

    Google Scholar 

  • Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylmide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Ann Biochem 112:195–203

    Google Scholar 

  • Colas B, Maroux S (1980) Simultaneous isolation of brush border and basolateral membrane from rabbit enterocytes. Presence of brush border hydrolases in the basolateral membrane of rabbit enterocytes. Biochim Biophys Acta 600:406–420

    Google Scholar 

  • Delovitch TH, Fegelman A, Barber BH, Frelinger JA (1979) Immunochemical characterisation of the Ly-8.2 murine lymphocyte alloantigen: possible relationship to actin. J Immunol 122:326–333

    Google Scholar 

  • Dunham ET, Glynn IM (1961) Adenosine-triphosphatase activity and the active movements of alkali metal ions. J Physiol 156:174–283

    Google Scholar 

  • Fambrough DM, Baynes EK (1983) Multiple forms of (Na+ + K+)-ATPase in the chicken. Selective detection of the major nerve, skeletal muscle, and kidney form by a monoclonal antibody. J Biol Chem 258:3926–3935

    Google Scholar 

  • Fujita M, Ohta H, Kawai K, Matsui H, Nakao M (1972) Differential isolation of microvillous and basolateral plasma membrane from intestinal mucosa: mutually exclusive distribution of digestive enzymes and ouabain-sensitive ATPase. Biochim Biophys Acta 274:336–347

    Google Scholar 

  • Goding JW (1976) Conjugation of antibodies with fluorochromes: modification to the standard methods. J Immunol 13:215–226

    Google Scholar 

  • Golstein P, Goridis C, Schmitt-Verhulst AM, Hayot B, Pierres A, van Agthoven A, Kaufmann Y, Eshhar Z, Pierres M (1983) Lymphoid cell surface interaction structures detected using cytolysis-inhibiting monoclonal antibodies. Immunol Rev 68:5–42

    Google Scholar 

  • Goridis C, Joher AM, Hirsch M, Schachner M (1978) Cell surface proteins of cultured brain cells and their recognition by anticerebellum (anti-NS-4) antiserum. J Neurochem 31:531–539

    Google Scholar 

  • Gorvel JP, Liabeuf A, Massey D, Liot D, Goridis C, Maroux S (1983) Sodium- and potassium-dependent adenosinetriphosphatase recognition on mouse organs by means of a monoclonal antibody. Cell Tissue Res 234:619–632

    Google Scholar 

  • Hart DA (1981) Evidence that lithium ions can modulate lectin stimulation of lymphoid cells by multiple mechanisms. Cell Immunol 58:372–384

    Google Scholar 

  • Hirn M, Pierres M, Deagostini-Bazin H, Hirsch M, Goridis C (1981) Monoclonal antibody against cell surface glycoprotein of neurons. Brain Res 214:433–439

    Google Scholar 

  • Hirn M, Pierres M, Deagostini-Bazin H, Hirsch MR, Goridis C, Ghandour MS, Langley OK, Gombos G (1982) A new brain cell surface glycoprotein identified by monoclonal antibody. Neuroscience 7:239–250

    Google Scholar 

  • Hirn M, Ghandour MS, Deagostini-Bazin H, Goridis C (1983) Molecular heterogeneity and structural evolution during cerebellar ontogeny detected by monoclonal antibody of the mouse cell surface antigen BSP-2. Brain Res 265:87–100

    Google Scholar 

  • Hobbs A, Albers RW (1980) The structure of proteins involved in active membrane transport. Annu Rev Biophys Bioeng 9:259–291

    Google Scholar 

  • Hokin LE, Dahl JL, Deupree JD, Dixon JR, Hackney JF, Perdue JF (1973) Characterization of the sodium-potassium transport adenosinetriphosphatase. J Biol Chem 248:2593–2605

    Google Scholar 

  • Hubbard AL, Cohn ZA (1972) The iodination of the red cell membrane. J Cell Biol 55:390–405

    Google Scholar 

  • Jay G, Palladino MA, Khoury G, Old LJ (1982) Mouse Lyt-2 antigen: Evidence for two heterodimers with a common subunit. Proc Natl Acad Sci (USA) 79:2654–2657

    Google Scholar 

  • Jones PP (1977) Analysis of H-2 and Ia molecules by two dimensional gel electrophoresis. J Exp Med 146:1261–1279

    Google Scholar 

  • Jorgensen PL (1982) Mechanisms of the Na+, K+ pump. Protein structure and conformations of the pure (Na++K+)-ATPase. Biochim Biophys Acta 694:27–68

    Google Scholar 

  • Jorgensen PL, Karlish SJD, Gitler C (1982) Evidence for the organization of the transmembrane segments of (Na, K)-ATPase based on labeling lipid-embedded and surface domains of the α-subunit. J Biol Chem 257:7435–7442

    Google Scholar 

  • Koch KS, Leffert HL (1979) Increased sodium ion influx is necessary to initiate rat hepatocyte proliferation. Cell 18:153–163

    Google Scholar 

  • Kovářů H, Kovářů F (1979) Role of surface enzymes in mitogenic and allogenic interactions of brain cortex or spleen cells of mice. Simple and high yield isolation of spleen lymphocytes. Folia Biol (Praha) 25:415–416

    Google Scholar 

  • Kyte J (1972) Properties of the two polypeptides of sodium- and potassium-dependent adenosine triphosphatase. J Biol Chem 247:7642–7649

    Google Scholar 

  • Kyte J (1976a) Immunoferritin determination of the distribution of (Na+ + K+)-ATPase over the plasma membranes of renal convoluted tubules. I. Distal segment. J Cell Biol 68:287–303

    Google Scholar 

  • Kyte J (1976b) Immunoferritin determination of the distribution of (Na+ +K+)-ATPase over the plasma membranes of renal convoluted tubules. II. Proximal segment. J Cell Biol 68:304–318

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227:680–685

    Google Scholar 

  • Ledbetter JA, Rouse RV, Spedding Micklem H, Herzenberg LA (1980) T cell subsets defined by expression of Lyt-1,2,3 and Thy-1 antigens. Two parameter immunofluorescence and cytotoxicity analysis with monoclonal antibodies modify current viewt. J Exp Med 152:280–295

    Google Scholar 

  • Ledbetter JA, Seaman WE, Tsu TT, Herzenberg LA (1981) Lyt-2 and Lyt-3 antigens are on two different polypeptide subunits linked by disulfide bonds. Relationship of subunits to T cell cytolytic activity. J Exp Med 153:1503–1516

    Google Scholar 

  • Mendoza SA, Wigglesworth NH, Pohjanpelto P, Rozengurt E (1980) Na entry and Na-K pump activity in murine, hamster and human cells. J Cell Physiol 103:17–27

    Google Scholar 

  • Moolenaar WH, Mummery CL, Van der Saag PT, de Laat SW (1981) Rapid ionic events and the initiation of growth in serum stimulated neuroblastoma cells. Cell 23:789–798

    Google Scholar 

  • Moolenaar WH, Yarden Y, de Laat SW, Schlessinger J (1982) Epidermal growth factor induces electrically silent Na+ influx in human fibroblasts. J Biol Chem 257:8502–8506

    Google Scholar 

  • Pierres M, Goridis C, Golstein P (1982) Inhibition of murine T cell-mediated cytolysis and T cell proliferation by a rat monoclonal antibody immunoprecipitating two lymphoid cell surface polypeptides of 94000 and 180000 molecular weight. Eur J Immunol 12:60–69

    Google Scholar 

  • Quastel MR, Kaplan JG (1968) Inhibition by ouabain of human lymphocyte transformation induced by phytohaemagglutinin in vitro. Nature (Lond) 219:198–200

    Google Scholar 

  • Quastel MR, Kaplan JG (1970a) Early stimulation of potassium uptake in lymphocyte treated with PHA. Exp Cell Res 63:230–233

    Google Scholar 

  • Quastel MR, Kaplan JG (1970b) Lymphocyte stimulation: the effect of ouabain on nucleic acid and protein synthesis. Exp Cell Res 62:407–420

    Google Scholar 

  • Raff MC (1971) Surface antigenic markers to distinguish T and B lymphocytes in mice. Transplant Rev 6:52–80

    Google Scholar 

  • Reggio H, Coudrier E, Louvard D (1982) Surface and cytoplasmic domains in polarized epithelial cells. In: Hoffman JF, Giebisch GH, Bolis L (eds) Membranes in growth and development. Alan R Liss, New York, pp 89–105

    Google Scholar 

  • Rode HN, Mähler B, Loracher A, Resch K (1979) Functional mosaicism of the lymphocyte plasma membrane. II. Characterization of membrane subfractions of activated thymocytes. Eur J Immunol 9:402–408

    Google Scholar 

  • Rothenberg P, Glaser L, Schlesinger P, Cassel D (1983) Epidermal growth factor stimulates amiloride-sensitive 22Na+ uptake in A431 cells. J Biol Chem 258:4883–4889

    Google Scholar 

  • Sarmiento M, Loken MR, Trowbridge IS, Coffman RL, Fitch FW (1982) High molecular weight lymphocyte surface proteins are structurally related and are expressed on different cell populations at different times during lymphocyte maturation and differentiation. J Immunol 128:1676–1684

    Google Scholar 

  • Schenk DB, Leffert HL (1983) Monoclonal antibodies to rat Na+, K+-ATPase block enzymatic activity. Proc Natl Acad Sci (USA) 80:5281–5285

    Google Scholar 

  • Schwartz A, Lindmayer GE, Allen JC (1975) The sodium-potassium adenosine triphosphatase: pharmacological, physiological and biochemical aspects. Pharmacol Rev 27:3–124

    Google Scholar 

  • Segrest JP, Jackson RL (1973) Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Methods Enzymol 28:54–63

    Google Scholar 

  • Sharkey RG (1983) Lactoperoxidase-catalyzed iodination of sodium and potassium ion-activated adenosine triphosphatase in the madin-darby canine kidney epithelial cell line and canine renal membranes. Biochim Biophys Acta 730:327–341

    Google Scholar 

  • Smith JB, Rozengurt E (1978) Serum stimulates the Na+, K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci USA 75:5560–5564

    Google Scholar 

  • Specht SC, Sweadner KJ (1984) Two different Na, K-ATPases in the optic nerve: cells of origin and axonal transport. Proc Natl Acad Sci USA 81:1234–1238

    Google Scholar 

  • Sweadner KJ (1979) Two molecular forms of (Na++K+)-stimulated ATPase in brain. Separation, and difference in affinity for strophanthidin. J Biol Chem 254:6060–6067

    Google Scholar 

  • Sweadner KJ, Goldin SM (1975) Reconstitution of active ion tansport by the sodium and potassium ion-stimulated adenosine triphosphatase from canine brain. J Biol Chem 250:4022–4024

    Google Scholar 

  • Szamel M, Schneider S, Resch K (1981) Functional interrelationship between (Na++K+)-ATPase and lysolecithin acetyltransferase in plasma membranes of mitogen-stimulated rabbit thymocytes. J Biol Chem 256:9198–9204

    Google Scholar 

  • Wallick ET, Lane LK, Schwartz A (1979) Biochemical mechanism of the sodium pump. Ann Rev Physiol 41:397–411

    Google Scholar 

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Liabeuf, A., Gorvel, JP. & Goridis, C. Recognition of sodium- and potassium-dependent adenosine triphosphatase on mouse lymphoid cells by means of a monoclonal antibody. Cell Tissue Res. 238, 253–261 (1984). https://doi.org/10.1007/BF00217297

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