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. 1989 Dec 1;8(12):3759–3765. doi: 10.1002/j.1460-2075.1989.tb08552.x

Regulated expression of Mg2+ binding epitope on leukocyte integrin alpha subunits.

I Dransfield 1, N Hogg 1
PMCID: PMC402061  PMID: 2479549

Abstract

The leukocyte integrins LFA-1, CR3 and p150,95 are a family of heterodimeric receptors that mediate divalent cation-dependent cellular adhesion reactions. In this study we describe a novel antibody-defined epitope present on the leukocyte integrin alpha subunits indicating that the antibody recognizes a structural feature common to all three polypeptides. Antibody recognition further differs from that of previously described anti-leukocyte integrin antibodies in that it is strictly dependent upon the presence of Mg2+. This suggests that the epitope is located within, or in close proximity to, the three conserved cation binding domains and is therefore a measure of Mg2+ bound to the leukocyte integrins and thus reflects functionally active molecules. The epitope can be induced on polymorphonuclear leukocytes, a subset of T cells and on monocytes but is absent or much reduced at low temperature or in the presence of metabolic inhibitors. These observations have considerable implications for the regulation of leukocyte integrin function suggesting that Mg2+ binding to the extracellular domain(s) of the alpha subunits is controlled from within the cell. We suggest that one mechanism by which ligand binding by these molecules can be 'switched' on and off in response to external signals is by regulation of Mg2+ binding to these molecules, converting from the cation-free, 'inactive' to the Mg2+-bound, 'active' form.

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Selected References

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  1. Ackerman S. K., Douglas S. D. Purification of human monocytes on microexudate-coated surfaces. J Immunol. 1978 Apr;120(4):1372–1374. [PubMed] [Google Scholar]
  2. Altieri D. C., Edgington T. S. A monoclonal antibody reacting with distinct adhesion molecules defines a transition in the functional state of the receptor CD11b/CD18 (Mac-1). J Immunol. 1988 Oct 15;141(8):2656–2660. [PubMed] [Google Scholar]
  3. Anderson D. C., Springer T. A. Leukocyte adhesion deficiency: an inherited defect in the Mac-1, LFA-1, and p150,95 glycoproteins. Annu Rev Med. 1987;38:175–194. doi: 10.1146/annurev.me.38.020187.001135. [DOI] [PubMed] [Google Scholar]
  4. Buyon J. P., Abramson S. B., Philips M. R., Slade S. G., Ross G. D., Weissmann G., Winchester R. J. Dissociation between increased surface expression of gp165/95 and homotypic neutrophil aggregation. J Immunol. 1988 May 1;140(9):3156–3160. [PubMed] [Google Scholar]
  5. Chatila T. A., Geha R. S. Phosphorylation of T cell membrane proteins by activators of protein kinase C. J Immunol. 1988 Jun 15;140(12):4308–4314. [PubMed] [Google Scholar]
  6. Cheresh D. A., Pytela R., Pierschbacher M. D., Klier F. G., Ruoslahti E., Reisfeld R. A. An Arg-Gly-Asp-directed receptor on the surface of human melanoma cells exists in an divalent cation-dependent functional complex with the disialoganglioside GD2. J Cell Biol. 1987 Sep;105(3):1163–1173. doi: 10.1083/jcb.105.3.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Corbi A. L., Kishimoto T. K., Miller L. J., Springer T. A. The human leukocyte adhesion glycoprotein Mac-1 (complement receptor type 3, CD11b) alpha subunit. Cloning, primary structure, and relation to the integrins, von Willebrand factor and factor B. J Biol Chem. 1988 Sep 5;263(25):12403–12411. [PubMed] [Google Scholar]
  8. Corbi A. L., Miller L. J., O'Connor K., Larson R. S., Springer T. A. cDNA cloning and complete primary structure of the alpha subunit of a leukocyte adhesion glycoprotein, p150,95. EMBO J. 1987 Dec 20;6(13):4023–4028. doi: 10.1002/j.1460-2075.1987.tb02746.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dooley D. C., Simpson J. F., Meryman H. T. Isolation of large numbers of fully viable human neutrophils: a preparative technique using percoll density gradient centrifugation. Exp Hematol. 1982 Aug;10(7):591–599. [PubMed] [Google Scholar]
  10. Dougherty G. J., Dransfield I., Hogg N. Identification of a novel monocyte cell surface molecule involved in the generation of antigen-induced proliferative responses. Eur J Immunol. 1988 Dec;18(12):2067–2071. doi: 10.1002/eji.1830181229. [DOI] [PubMed] [Google Scholar]
  11. Ey P. L., Prowse S. J., Jenkin C. R. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  12. Gailit J., Ruoslahti E. Regulation of the fibronectin receptor affinity by divalent cations. J Biol Chem. 1988 Sep 15;263(26):12927–12932. [PubMed] [Google Scholar]
  13. Hemler M. E. Adhesive protein receptors on hematopoietic cells. Immunol Today. 1988 Apr;9(4):109–113. doi: 10.1016/0167-5699(88)91280-7. [DOI] [PubMed] [Google Scholar]
  14. Hildreth J. E., August J. T. The human lymphocyte function-associated (HLFA) antigen and a related macrophage differentiation antigen (HMac-1): functional effects of subunit-specific monoclonal antibodies. J Immunol. 1985 May;134(5):3272–3280. [PubMed] [Google Scholar]
  15. Hildreth J. E., Gotch F. M., Hildreth P. D., McMichael A. J. A human lymphocyte-associated antigen involved in cell-mediated lympholysis. Eur J Immunol. 1983 Mar;13(3):202–208. doi: 10.1002/eji.1830130305. [DOI] [PubMed] [Google Scholar]
  16. Hogg N., Selvendran Y. An anti-human monocyte/macrophage monoclonal antibody, reacting most strongly with macrophages in lymphoid tissue. Cell Immunol. 1985 May;92(2):247–253. doi: 10.1016/0008-8749(85)90006-1. [DOI] [PubMed] [Google Scholar]
  17. Hogg N., Takacs L., Palmer D. G., Selvendran Y., Allen C. The p150,95 molecule is a marker of human mononuclear phagocytes: comparison with expression of class II molecules. Eur J Immunol. 1986 Mar;16(3):240–248. doi: 10.1002/eji.1830160306. [DOI] [PubMed] [Google Scholar]
  18. Hogg N. The leukocyte integrins. Immunol Today. 1989 Apr;10(4):111–114. doi: 10.1016/0167-5699(89)90238-7. [DOI] [PubMed] [Google Scholar]
  19. Hubbard A. L., Cohn Z. A. Externally disposed plasma membrane proteins. I. Enzymatic iodination of mouse L cells. J Cell Biol. 1975 Feb;64(2):438–460. doi: 10.1083/jcb.64.2.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  21. Ignatius M. J., Reichardt L. F. Identification of a neuronal laminin receptor: an Mr 200K/120K integrin heterodimer that binds laminin in a divalent cation-dependent manner. Neuron. 1988 Oct;1(8):713–725. doi: 10.1016/0896-6273(88)90170-5. [DOI] [PubMed] [Google Scholar]
  22. Julius M. H., Simpson E., Herzenberg L. A. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
  23. Larson R. S., Corbi A. L., Berman L., Springer T. Primary structure of the leukocyte function-associated molecule-1 alpha subunit: an integrin with an embedded domain defining a protein superfamily. J Cell Biol. 1989 Feb;108(2):703–712. doi: 10.1083/jcb.108.2.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Malhotra V., Hogg N., Sim R. B. Ligand binding by the p150,95 antigen of U937 monocytic cells: properties in common with complement receptor type 3 (CR3). Eur J Immunol. 1986 Sep;16(9):1117–1123. doi: 10.1002/eji.1830160915. [DOI] [PubMed] [Google Scholar]
  25. Marlin S. D., Springer T. A. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1). Cell. 1987 Dec 4;51(5):813–819. doi: 10.1016/0092-8674(87)90104-8. [DOI] [PubMed] [Google Scholar]
  26. Martz E. Immune T lymphocyte to tumor cell adhesion. Magnesium sufficient, calcium insufficient. J Cell Biol. 1980 Mar;84(3):584–598. doi: 10.1083/jcb.84.3.584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Martz E. LFA-1 and other accessory molecules functioning in adhesions of T and B lymphocytes. Hum Immunol. 1987 Jan;18(1):3–37. doi: 10.1016/0198-8859(87)90110-8. [DOI] [PubMed] [Google Scholar]
  28. Miller L. J., Bainton D. F., Borregaard N., Springer T. A. Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. J Clin Invest. 1987 Aug;80(2):535–544. doi: 10.1172/JCI113102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Patarroyo M., Beatty P. G., Fabre J. W., Gahmberg C. G. Identification of a cell surface protein complex mediating phorbol ester-induced adhesion (binding) among human mononuclear leukocytes. Scand J Immunol. 1985 Aug;22(2):171–182. doi: 10.1111/j.1365-3083.1985.tb01869.x. [DOI] [PubMed] [Google Scholar]
  30. Pytela R. Amino acid sequence of the murine Mac-1 alpha chain reveals homology with the integrin family and an additional domain related to von Willebrand factor. EMBO J. 1988 May;7(5):1371–1378. doi: 10.1002/j.1460-2075.1988.tb02953.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ross G. D., Newman S. L., Lambris J. D., Devery-Pocius J. E., Cain J. A., Lachmann P. J. Generation of three different fragments of bound C3 with purified factor I or serum. II. Location of binding sites in the C3 fragments for factors B and H, complement receptors, and bovine conglutinin. J Exp Med. 1983 Aug 1;158(2):334–352. doi: 10.1084/jem.158.2.334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sanchez-Madrid F., Nagy J. A., Robbins E., Simon P., Springer T. A. A human leukocyte differentiation antigen family with distinct alpha-subunits and a common beta-subunit: the lymphocyte function-associated antigen (LFA-1), the C3bi complement receptor (OKM1/Mac-1), and the p150,95 molecule. J Exp Med. 1983 Dec 1;158(6):1785–1803. doi: 10.1084/jem.158.6.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shaw S., Luce G. E., Quinones R., Gress R. E., Springer T. A., Sanders M. E. Two antigen-independent adhesion pathways used by human cytotoxic T-cell clones. Nature. 1986 Sep 18;323(6085):262–264. doi: 10.1038/323262a0. [DOI] [PubMed] [Google Scholar]
  34. Simmons D., Makgoba M. W., Seed B. ICAM, an adhesion ligand of LFA-1, is homologous to the neural cell adhesion molecule NCAM. Nature. 1988 Feb 18;331(6157):624–627. doi: 10.1038/331624a0. [DOI] [PubMed] [Google Scholar]
  35. Smith C. W., Marlin S. D., Rothlein R., Toman C., Anderson D. C. Cooperative interactions of LFA-1 and Mac-1 with intercellular adhesion molecule-1 in facilitating adherence and transendothelial migration of human neutrophils in vitro. J Clin Invest. 1989 Jun;83(6):2008–2017. doi: 10.1172/JCI114111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Springer T. A., Dustin M. L., Kishimoto T. K., Marlin S. D. The lymphocyte function-associated LFA-1, CD2, and LFA-3 molecules: cell adhesion receptors of the immune system. Annu Rev Immunol. 1987;5:223–252. doi: 10.1146/annurev.iy.05.040187.001255. [DOI] [PubMed] [Google Scholar]
  37. Staatz W. D., Rajpara S. M., Wayner E. A., Carter W. G., Santoro S. A. The membrane glycoprotein Ia-IIa (VLA-2) complex mediates the Mg++-dependent adhesion of platelets to collagen. J Cell Biol. 1989 May;108(5):1917–1924. doi: 10.1083/jcb.108.5.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Staunton D. E., Dustin M. L., Springer T. A. Functional cloning of ICAM-2, a cell adhesion ligand for LFA-1 homologous to ICAM-1. Nature. 1989 May 4;339(6219):61–64. doi: 10.1038/339061a0. [DOI] [PubMed] [Google Scholar]
  39. Todd R. F., 3rd, Arnaout M. A., Rosin R. E., Crowley C. A., Peters W. A., Babior B. M. Subcellular localization of the large subunit of Mo1 (Mo1 alpha; formerly gp 110), a surface glycoprotein associated with neutrophil adhesion. J Clin Invest. 1984 Oct;74(4):1280–1290. doi: 10.1172/JCI111538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wright S. D., Detmers P. A., Jong M. T., Meyer B. C. Interferon-gamma depresses binding of ligand by C3b and C3bi receptors on cultured human monocytes, an effect reversed by fibronectin. J Exp Med. 1986 May 1;163(5):1245–1259. doi: 10.1084/jem.163.5.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wright S. D., Rao P. E., Van Voorhis W. C., Craigmyle L. S., Iida K., Talle M. A., Westberg E. F., Goldstein G., Silverstein S. C. Identification of the C3bi receptor of human monocytes and macrophages by using monoclonal antibodies. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5699–5703. doi: 10.1073/pnas.80.18.5699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wright S. D., Silverstein S. C. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes. J Exp Med. 1982 Oct 1;156(4):1149–1164. doi: 10.1084/jem.156.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wright S. D., Weitz J. I., Huang A. J., Levin S. M., Silverstein S. C., Loike J. D. Complement receptor type three (CD11b/CD18) of human polymorphonuclear leukocytes recognizes fibrinogen. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7734–7738. doi: 10.1073/pnas.85.20.7734. [DOI] [PMC free article] [PubMed] [Google Scholar]

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