Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1989 Sep;84(3):793–801. doi: 10.1172/JCI114238

Role of catalytic and lysine-binding sites in plasmin-induced neutrophil adherence to endothelium.

S K Lo 1, T J Ryan 1, N Gilboa 1, L Lai 1, A B Malik 1
PMCID: PMC329721  PMID: 2569479

Abstract

Plasmin resulted in increased neutrophil adherence to cultured ovine pulmonary artery endothelial cell monolayers in a concentration-dependent manner (10(-12)-10(-7) M). The adherence response increased fivefold above baseline within 60 min after addition of plasmin (10(-8) M) and the response persisted up to 30 min after removal of plasmin. The neutrophil adherence was mediated by the action of plasmin on neutrophils rather than endothelial cells. The response was the result of an increase in functional activity of CD18 neutrophil cell surface adhesive glycoprotein. Neutrophil adherence was inhibited by pretreatment of neutrophils with MAbs IB4 and 60.3 targeted against the beta chain of the CD18, whereas control isotypic MAb 60.5 against HLA class I antigen had no effect. The plasmin catalytic site was not involved in the response. Lys-plasminogen had reduced adherence-promoting activity relative to plasmin, whereas glu-plasminogen had no effect. Elastase-derived plasminogen fragments corresponding to kringle 1+2+3 and kringle 4 (both of which contained the lysine-binding sites) possessed neutrophil adherence-promoting activities similar to plasmin, whereas miniplasminogen (which contains the catalytic site but no lysine-binding sites) had minimal effect, indicating the involvement of lysine-binding sites in the response. Blocking lysine-binding sites of plasmin and elastase-derived plasminogen fragments with tranexamic acid (IC50 of 5 mM) inhibited neutrophil adherence. A monospecific polyclonal antibody against the lysine-binding sites also reduced the neutrophil adherence-promoting activity of plasmin. The results indicate that plasmin induces neutrophil adherence to the endothelium and that the effect is mediated by lysine-binding sites on plasmin.

Full text

PDF
793

Images in this article

796Image
on p.796
797Image
on p.797

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. 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]
  2. Bauer P. I., Machovich R., Büki K. G., Csonka E., Koch S. A., Horváth I. Interaction of plasmin with endothelial cells. Biochem J. 1984 Feb 15;218(1):119–124. doi: 10.1042/bj2180119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bevilacqua M. P., Pober J. S., Wheeler M. E., Cotran R. S., Gimbrone M. A., Jr Interleukin 1 acts on cultured human vascular endothelium to increase the adhesion of polymorphonuclear leukocytes, monocytes, and related leukocyte cell lines. J Clin Invest. 1985 Nov;76(5):2003–2011. doi: 10.1172/JCI112200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bizios R., Lai L. C., Cooper J. A., Del Vecchio P. J., Malik A. B. Thrombin-induced adherence of neutrophils to cultured endothelial monolayers: increased endothelial adhesiveness. J Cell Physiol. 1988 Feb;134(2):275–280. doi: 10.1002/jcp.1041340214. [DOI] [PubMed] [Google Scholar]
  5. Collen D. Molecular mechanisms of fibrinolysis and their application to fibrin-specific thrombolytic therapy. J Cell Biochem. 1987 Feb;33(2):77–86. doi: 10.1002/jcb.240330202. [DOI] [PubMed] [Google Scholar]
  6. Cooper J. A., Lo S. K., Malik A. B. Fibrin is a determinant of neutrophil sequestration in the lung. Circ Res. 1988 Oct;63(4):735–741. doi: 10.1161/01.res.63.4.735. [DOI] [PubMed] [Google Scholar]
  7. Cooper J. A., Solano S. J., Bizios R., Kaplan J. E., Malik A. B. Pulmonary neutrophil kinetics after thrombin-induced intravascular coagulation. J Appl Physiol Respir Environ Exerc Physiol. 1984 Sep;57(3):826–832. doi: 10.1152/jappl.1984.57.3.826. [DOI] [PubMed] [Google Scholar]
  8. Del Vecchio P. J., Bizios R., Holleran L. A., Judge T. K., Pinto G. L. Inhibition of human scleral fibroblast proliferation with heparin. Invest Ophthalmol Vis Sci. 1988 Aug;29(8):1272–1276. [PubMed] [Google Scholar]
  9. Detmers P. A., Wright S. D., Olsen E., Kimball B., Cohn Z. A. Aggregation of complement receptors on human neutrophils in the absence of ligand. J Cell Biol. 1987 Sep;105(3):1137–1145. doi: 10.1083/jcb.105.3.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gilboa N., Kaplan J. E. Plasma fibronectin enhances fibrinolytic system in vitro. Thromb Haemost. 1985 Oct 30;54(3):639–644. [PubMed] [Google Scholar]
  11. Gimbrone M. A., Jr, Brock A. F., Schafer A. I. Leukotriene B4 stimulates polymorphonuclear leukocyte adhesion to cultured vascular endothelial cells. J Clin Invest. 1984 Oct;74(4):1552–1555. doi: 10.1172/JCI111570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guccione M. A., Kinlough-Rathbone R. L., Packham M. A., Harfenist E. J., Rand M. L., Greenberg J. P., Perry D. W., Mustard J. F. Effects of plasmin on rabbit platelets. Thromb Haemost. 1985 Feb 18;53(1):8–14. [PubMed] [Google Scholar]
  13. Hajjar K. A., Harpel P. C., Jaffe E. A., Nachman R. L. Binding of plasminogen to cultured human endothelial cells. J Biol Chem. 1986 Sep 5;261(25):11656–11662. [PubMed] [Google Scholar]
  14. Harlan J. M., Killen P. D., Senecal F. M., Schwartz B. R., Yee E. K., Taylor R. F., Beatty P. G., Price T. H., Ochs H. D. The role of neutrophil membrane glycoprotein GP-150 in neutrophil adherence to endothelium in vitro. Blood. 1985 Jul;66(1):167–178. [PubMed] [Google Scholar]
  15. Lerch P. G., Rickli E. E., Lergier W., Gillessen D. Localization of individual lysine-binding regions in human plasminogen and investigations on their complex-forming properties. Eur J Biochem. 1980;107(1):7–13. doi: 10.1111/j.1432-1033.1980.tb04617.x. [DOI] [PubMed] [Google Scholar]
  16. Lucas M. A., Fretto L. J., McKee P. A. The binding of human plasminogen to fibrin and fibrinogen. J Biol Chem. 1983 Apr 10;258(7):4249–4256. [PubMed] [Google Scholar]
  17. Malik A. B. Pulmonary microembolism. Physiol Rev. 1983 Jul;63(3):1114–1207. doi: 10.1152/physrev.1983.63.3.1114. [DOI] [PubMed] [Google Scholar]
  18. Markus G., DePasquale J. L., Wissler F. C. Quantitative determination of the binding of epsilon-aminocaproic acid to native plasminogen. J Biol Chem. 1978 Feb 10;253(3):727–732. [PubMed] [Google Scholar]
  19. Miles L. A., Ginsberg M. H., White J. G., Plow E. F. Plasminogen interacts with human platelets through two distinct mechanisms. J Clin Invest. 1986 Jun;77(6):2001–2009. doi: 10.1172/JCI112529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miles L. A., Plow E. F. Binding and activation of plasminogen on the platelet surface. J Biol Chem. 1985 Apr 10;260(7):4303–4311. [PubMed] [Google Scholar]
  21. Miles L. A., Plow E. F. Receptor mediated binding of the fibrinolytic components, plasminogen and urokinase, to peripheral blood cells. Thromb Haemost. 1987 Oct 28;58(3):936–942. [PubMed] [Google Scholar]
  22. Miles L. A., Plow E. F. Topography of the high-affinity lysine binding site of plasminogen as defined with a specific antibody probe. Biochemistry. 1986 Nov 4;25(22):6926–6933. doi: 10.1021/bi00370a028. [DOI] [PubMed] [Google Scholar]
  23. Niewiarowski S., Senyi A. F., Gillies P. Plasmin-induced platelet aggregation and platelet release reaction. Effects on hemostasis. J Clin Invest. 1973 Jul;52(7):1647–1659. doi: 10.1172/JCI107345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Plow E. F., Collen D. Immunochemical characterization of a low affinity lysine binding site within plasminogen. J Biol Chem. 1981 Nov 10;256(21):10864–10869. [PubMed] [Google Scholar]
  25. Plow E. F., Freaney D. E., Plescia J., Miles L. A. The plasminogen system and cell surfaces: evidence for plasminogen and urokinase receptors on the same cell type. J Cell Biol. 1986 Dec;103(6 Pt 1):2411–2420. doi: 10.1083/jcb.103.6.2411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pohlman T. H., Stanness K. A., Beatty P. G., Ochs H. D., Harlan J. M. An endothelial cell surface factor(s) induced in vitro by lipopolysaccharide, interleukin 1, and tumor necrosis factor-alpha increases neutrophil adherence by a CDw18-dependent mechanism. J Immunol. 1986 Jun 15;136(12):4548–4553. [PubMed] [Google Scholar]
  27. Ryan T. J., Keegan M. C. Photoaffinity labeling of functionally different lysine-binding sites in human plasminogen and plasmin. Biochim Biophys Acta. 1985 Aug 8;830(2):187–194. doi: 10.1016/0167-4838(85)90027-5. [DOI] [PubMed] [Google Scholar]
  28. Ryan T. J. Photoaffinity labeling of human plasmin and plasminogen. Pharmacol Ther. 1987;34(2):335–348. doi: 10.1016/0163-7258(87)90017-9. [DOI] [PubMed] [Google Scholar]
  29. Schafer A. I., Adelman B. Plasmin inhibition of platelet function and of arachidonic acid metabolism. J Clin Invest. 1985 Feb;75(2):456–461. doi: 10.1172/JCI111720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schafer A. I., Maas A. K., Ware J. A., Johnson P. C., Rittenhouse S. E., Salzman E. W. Platelet protein phosphorylation, elevation of cytosolic calcium, and inositol phospholipid breakdown in platelet activation induced by plasmin. J Clin Invest. 1986 Jul;78(1):73–79. doi: 10.1172/JCI112576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schleimer R. P., Rutledge B. K. Cultured human vascular endothelial cells acquire adhesiveness for neutrophils after stimulation with interleukin 1, endotoxin, and tumor-promoting phorbol diesters. J Immunol. 1986 Jan;136(2):649–654. [PubMed] [Google Scholar]
  32. 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]
  33. Tonnesen M. G., Smedly L. A., Henson P. M. Neutrophil-endothelial cell interactions. Modulation of neutrophil adhesiveness induced by complement fragments C5a and C5a des arg and formyl-methionyl-leucyl-phenylalanine in vitro. J Clin Invest. 1984 Nov;74(5):1581–1592. doi: 10.1172/JCI111574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vedder N. B., Harlan J. M. Increased surface expression of CD11b/CD18 (Mac-1) is not required for stimulated neutrophil adherence to cultured endothelium. J Clin Invest. 1988 Mar;81(3):676–682. doi: 10.1172/JCI113372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Váli Z., Patthy L. Location of the intermediate and high affinity omega-aminocarboxylic acid-binding sites in human plasminogen. J Biol Chem. 1982 Feb 25;257(4):2104–2110. [PubMed] [Google Scholar]
  36. Wright S. D., Meyer B. C. Phorbol esters cause sequential activation and deactivation of complement receptors on polymorphonuclear leukocytes. J Immunol. 1986 Mar 1;136(5):1759–1764. [PubMed] [Google Scholar]
  37. 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]
  38. Zimmerman G. A., McIntyre T. M., Prescott S. M. Thrombin stimulates the adherence of neutrophils to human endothelial cells in vitro. J Clin Invest. 1985 Dec;76(6):2235–2246. doi: 10.1172/JCI112232. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES