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Protein 1a: A major wheat germ agglutinin binding protein on the surface of human granulocytes associated with the cytoskeleton

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Abstract

Lectin-receptors on leukocyte and endothelial surfaces are becoming more important in the light of increasing evidence which implicates lectin-carbohydrate interactions in diverse physiological phenomena. This study reports the identification of a major 118 kDa granulocyte surface protein, (Protein 1a) which binds the lectin wheat germ agglutinin (WGA), and is distinctly different from reported WGA binding granulocyte membrane proteins. Protein 1a has been isolated from the Triton-soluble and Triton-insoluble lysates of normal individuals and patients with Chronic Myeloid Leukemia (CML) using a combination of differential solubilization, lectin affinity, ion exchange chromatography and HPLC. The protein from the detergent lysates of both normal and CML granulocytes has similar pI values, lectin affinities, and hydrophobicity. However, its solubility in Triton is different in the two cell types. In 71% of CML cases examined, Protein 1a exhibits decreased Triton solubility suggesting its increased association with the cytoskeleton (CSK). Stimulation of normal granulocytes with WGA leads to the translocation of the soluble form of Protein 1a to the Triton-insoluble fraction. This cytoskeletal recruitment of Protein 1a is sustained only under conditions of excess WGA and occupied receptor. The CSK disruptive agent dihydrocytochalasin B (H2CB) releases the insoluble form of the receptor into the Triton-soluble fraction. Investigation of a CSK-involving process such as ligand internalization revealed that CML granulocytes exhibit slower kinetics of internalization of fluorescent WGA molecules. Since Protein 1a is a major WGA receptor on the granulocyte surface, its decreased Triton solubility in CML granulocytes suggests that this may be one of the factors contributing to the defective receptor-mediated endocytosis of WGA by CML cells, arising as a consequence of altered membrane-CSK interaction — a nodal point in the signal transduction cascade.

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References

  1. Sharon N: Bacterial lectins, cell-cell recognition and infectious disease. FEBS Lett 217: 145–157, 1987

    PubMed  Google Scholar 

  2. Stoolman LM, Tenforde TS, Rosen SD: Phosphomannosyl receptors may participate in the adhesive interactions between lymphocytes and high endothelial venules. J Cell Biol 99: 1535–1540, 1984

    PubMed  Google Scholar 

  3. Bevilacqua MP: Endothelial-leukocyte adhesion molecules. Annu Rev Immunol 11: 767–804, 1993

    PubMed  Google Scholar 

  4. Moore KL, Varki A, McEver RP: GMP-140 binds to a glycoprotein receptor on human neutrophils: evidence for a lectin-like interaction. J Cell Biol 112: 491–499, 1991

    PubMed  Google Scholar 

  5. Springer TA: Traffic signals for lymphocyte recirculation and leukocyte emigration: The multistep paradigm. Cell 76: 301–314, 1994

    PubMed  Google Scholar 

  6. Christiansen NP, Skubitz KM: Identification of the major lectin-binding surface proteins of human neutrophils and alveolar macrophages. Blood 71: 1624–1632, 1988

    PubMed  Google Scholar 

  7. Kimberly RP, Tappe NJ, Merriam LT, Redecha PB, Edberg JC, Schwartzman S, Valinsky JE: Carbohydrates on human Fcγ receptors: Interdependence of the classical IgG and nonclassical lectin-binding sites on human FcγRIII expressed on neutrophils. J Immunol 142: 3923–3930, 1989

    PubMed  Google Scholar 

  8. Grob PM, David E, Warren TC, DeLeon RP, Farina PR, Homon CA: Characterization of a receptor for human monocyte-derived neutrophil chemotactic factor/Interleukin-8. J Biol Chem 265: 8311–8316, 1990

    PubMed  Google Scholar 

  9. Allen RA, Traynor AE, Omann GM, Jesaitis AJ: The chemotactic peptide receptor — a model for future understanding of chemotactic disorders. Haematol Oncol Clinics of North America 2: 33–59, 1988

    Google Scholar 

  10. Suchard SJ, Boxer LA: Characterization and cytoskeletal association of a major cell surface glycoprotein, GP-140, in human neutrophils. J Clin Invest 84: 484–493, 1989

    PubMed  Google Scholar 

  11. Newman RA, Sutherland R, Greaves MF: The biochemical characterization of a cell surface antigen associated with acute lymphoblastic leukemia and lymphocyte precursors. J Immunol 126: 2024–2030, 1981

    PubMed  Google Scholar 

  12. Horejsi V, Bazil V: Surface proteins and glycoproteins of human leucocytes. Biochem J 253: 1–26, 1988

    PubMed  Google Scholar 

  13. Carlsson SR, Fukuda M: Isolation and characterization of leukosialin, a major sialoglycoprotein on human leukocytes. J Biol Chem 261: 12779–12786, 1986

    PubMed  Google Scholar 

  14. Kuroki M, Matsuo Y, Kinugasa T, Matsuoka Y: Three different NCA species, CGM6/CD67, NCA-95 and NCA-90, are comprised in the major 90 to 100 KDa band of granulocyte NCA detectable upon SDS-polyacrylamide gel electrophoresis. Biochem Biophys Res Commun 182: 501–506, 1992

    PubMed  Google Scholar 

  15. Strife A, Clarkson B: Biology of Chronic Myeloid Leukemia: Is discordant maturation the primary defect? Sem Hematol 25: 1–19, 1988

    Google Scholar 

  16. Anklesaria PN, Advani SH, Bhisey AN: Defective chemotaxis and adherence in granulocytes from chronic myeloid leukemia (CML) patients. Leuk Res 9: 641–648, 1985

    PubMed  Google Scholar 

  17. Pederson B: Functional and biochemical phenotype in relation to cellular age of differentiated neutrophils in chronic myeloid leukemia. Br J Hematol 51: 339–344, 1982

    Google Scholar 

  18. Zingde SM, Anklesaria PN, Advani SH, Bhisey AN, Gothoskar BP: Differential endocytosis of fluorescene isothiocyanate-Concanavalin A by normal and chronic myeloid leukemic granulocytes. Blut 55: 81–88, 1987

    PubMed  Google Scholar 

  19. Shirsat NV, Zingde SM, Advani SH, Gothoskar BP: The receptor for IgG on human normal and chronic myeloid granulocytes. Functional and biochemical characterization. Cancer Biochem Biophys 10: 235–245, 1989

    PubMed  Google Scholar 

  20. Zingde SM, Advani SH, Gothoskar BP: Plasma membrane proteins from human normal and chronic myeloid leukemic granulocytes. Identification and partial characterization of the Concanavalin A-binding and detergent resistant proteins. Blut 55: 89–100, 1987

    PubMed  Google Scholar 

  21. Hingorani R, Zingde SM, Tankkar A, Advani SH, Gothoskar BP: A major concanavalin A binding cell surface protein from normal and leukemic granulocytes. Isolation and Characterization. Ann Hematol 65: 175–183, 1992

    PubMed  Google Scholar 

  22. Boyum A: Separation of blood leukocytes, granulocytes and lymphocytes. Tissue Antigens 4: 269–274, 1974

    PubMed  Google Scholar 

  23. Fraker PJ, Speck JC Jr: Protein and cell surface iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3α, 6α-diphenylglycouril. Biochem Biophys Res Commun 80: 849–857, 1978

    PubMed  Google Scholar 

  24. Laemmli UK: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  25. Blum H, Beier H, Gross HJ: Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8: 93–99, 1987

    Google Scholar 

  26. Peterson G: A simplification of the protein assay method of Lowryet al. which is more generally applicable. Anal Biochem 83: 346–356, 1977

    PubMed  Google Scholar 

  27. O'Farrell PH: High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021, 1975

    PubMed  Google Scholar 

  28. Barondes SH: Bifunctional properties of lectins: lectins redefined. Trends Biochem Sci 13: 480–482, 1988

    PubMed  Google Scholar 

  29. Gabius HJ: Detection and functions of mammalian lectins-with emphasis on membrane lectins. Biochim Biophys Acta 1071: 1–18, 1991

    PubMed  Google Scholar 

  30. Renold-O'Donnel E, Kenney DM, Parkman R, Caims L, Savag B, Rosen FS: Characterization of a human lymphocyte surface sialoglycoprotein that is defective in Wiskott-Aldrich syndrome. J Exp Med 159: 1705–1723, 1984

    PubMed  Google Scholar 

  31. Niggli V, Burger MM: Interaction of the cytoskeleton with the plasma membrane. J Mem Biol 100: 97–121, 1987

    Google Scholar 

  32. Jesaitis AJ, Naemura JR, Sklar LA, Cochrane CG, Painter RG: Rapid modulation of N-formyl chemotactic peptide receptors on the surface of the human granulocytes: Formation of high-affinity ligand-receptor complexes in transient association with cytoskeleton. J Cell Biol 98: 1378–1387, 1984

    PubMed  Google Scholar 

  33. Sklar LA: Ligand-receptor dynamics and signal amplification in the neutrophil. Adv Immunol 39: 95–143, 1986

    PubMed  Google Scholar 

  34. Miettinen HM, Matter K, Hunziker W, Rose JK, Mellman I: Fc receptor endocytosis is controlled by a cytoplasmic domain determinant that actively prevents coated pit localization. J Cell Biol 116: 875–888, 1992

    PubMed  Google Scholar 

  35. Graham IL, Gresham HD, Brown EJ: An immobile subset of plasma membrane CD11b/CD18 (Mac-1) is involved in phagocytosis of targets recognized by multiple receptors. J Immunol 142: 2352–2358, 1989

    PubMed  Google Scholar 

  36. Atlas SJ, Lin S: Dihydrocytochalasin B: Biological effects and binding to 3T3 cells. J Cell Biol 76: 360–370, 1978

    PubMed  Google Scholar 

  37. Goldman DW, Gifford LA, Marotti T, Koo CH, Goetzl EJ: Molecular and cellular properties of human polymorphonuclear leukocyte receptors for leukotriene B4 Federation Proc 46: 200–203, 1987

    Google Scholar 

  38. Jack RM, Ezzell RM, Hartwig J, Fearon DT: Differential interaction of the C3b//C4b receptor and MHC Class I with the cytoskeleton of human neutrophils. J Immunol 137: 3996–4003, 1986

    PubMed  Google Scholar 

  39. Bennet V: Immunoreactive forms of human erythroid ankyrin are present in diverse cells and tissues. Nature 281: 597–599, 1979

    PubMed  Google Scholar 

  40. Valerius NH, Stendahl O, Hartwig JH, Stossel TP: Distribution of actin binding protein and myosin in polymorphonuclear leucocytes during locomotion and phagocytosis. Cell 24: 195–202, 1981

    PubMed  Google Scholar 

  41. Stevenson KB, Clark RA, Nauseef WM: Fodrin and band 4.1 in a plasma-membrane associated fraction of human neutrophils. Blood 74: 2136–2143, 1989

    PubMed  Google Scholar 

  42. Niggli V, Jenni V: Actin associated proteins in human neutrophils: identification and reorganization upon cell activation. Eur J Cell Biol 49: 366–372, 1989

    PubMed  Google Scholar 

  43. Lomax KJ, Malech HL, Gallin JI: The molecular biology of selected phagocyte defects. Blood Revs 3: 94–104, 1989

    Google Scholar 

  44. Rosse WF: Phosphatidylinositol-linked proteins and paroxysmal nocturnal haemoglobinuria. Blood 75: 1595–1601, 1990

    PubMed  Google Scholar 

  45. Naik NR, Bhisey AN, Advani SH: Flow cytometric studies on actin polymerization in PMN cells from chronic myeloid leukemia (CML) patients. Leuk Res 14: 921–930, 1990

    PubMed  Google Scholar 

  46. Tarachandani A, Advani SH, Bhisey AN: Chronic myeloid leukemic granulocytes have lower amounts of cytoplasmic actin. Leuk Res 17: 833–838, 1993

    PubMed  Google Scholar 

  47. Perez HD, Ong R, Khanna K, Banda D, Goldstein IM: Wheat germ agglutinin specifically inhibits formyl peptide induced polymorphonuclear leucocyte chemotaxis. J Immunol 129: 2718–2724, 1982

    PubMed  Google Scholar 

  48. Ozaki Y, Iwata J, Ohashi T: Mechanism of neutrophil chemiluminescence induced by wheat germ agglutinin: Partial characterization of the antigens recognized by wheat germ agglutinin. Blood 64: 1094–1102, 1984

    PubMed  Google Scholar 

  49. Bryant RW, Granzow CA, Siegel MI, Egan RW, Billah MM: Wheat germ agglutinin and other selected lectins increase synthesis of decayaccelerating factor in human endothelial cells. J Immunol 147: 1856–1862, 1991

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Cancer Research Institute, Tata Memorial Centre, Parel, 400 012, Bombay, India

    Padmaja Mehta, Surekha Zingde, Hema Desai & Balwant Gothoskar

  2. Tata Memorial Hospital, Tata Memorial Centre, Parel, 400 012, Bombay, India

    Suresh Advani

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  1. Padmaja Mehta
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  2. Surekha Zingde
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  4. Hema Desai
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  5. Balwant Gothoskar
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Mehta, P., Zingde, S., Advani, S. et al. Protein 1a: A major wheat germ agglutinin binding protein on the surface of human granulocytes associated with the cytoskeleton. Mol Cell Biochem 144, 153–165 (1995). https://doi.org/10.1007/BF00944395

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  • DOI: https://doi.org/10.1007/BF00944395

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