Abstract
In an attempt to define the molecular basis of the adherence of Aspergillus fumigatus conidia to the host tissues, a step which might be mediated by the recognition of basement membrane laminin or fibrinogen, we analyzed the binding of these glycoproteins by flow cytometry and a microtiter plate adherence assay. Flow cytometry revealed that the binding of fluorescein isothiocyanate-labeled laminin to conidia was saturable and specific. Moreover, the ability of conidia to bind laminin increased with their maturation. Competition experiments showed a cross-reactivity between laminin and fibrinogen binding and a lack of interactions with glycosaminoglycans. In addition, the binding of laminin was not inhibited by the different adhesive synthetic peptides tested. Furthermore, the microtiter plate assay of adherence to chymotrypsin degradation products of laminin or fibrinogen purified by gel filtration suggested a unique binding site common to sequential degradation fragments or the presence of multiple binding sites on the two ligands. Therefore, the role of carbohydrates in the recognition process was investigated. Among the carbohydrates tested, constitutive of the conidial wall or of the oligosaccharide side chains of laminin and fibrinogen, only N-acetylneuraminic acid and sialyllactose inhibited the binding of these glycoproteins to conidia. In conclusion, these results strengthen the idea that the laminin and fibrinogen receptors in A. fumigatus are identical and suggest an interaction mediated by a sialic acid-specific lectin of the conidial wall.
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- Annaix V., Bouchara J. P., Larcher G., Chabasse D., Tronchin G. Specific binding of human fibrinogen fragment D to Aspergillus fumigatus conidia. Infect Immun. 1992 May;60(5):1747–1755. doi: 10.1128/iai.60.5.1747-1755.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Annaix V., Bouchara J. P., Tronchin G., Senet J. M., Robert R. Structures involved in the binding of human fibrinogen to Candida albicans germ tubes. FEMS Microbiol Immunol. 1990 Oct;2(3):147–153. doi: 10.1111/j.1574-6968.1990.tb03514.x. [DOI] [PubMed] [Google Scholar]
- Beck K., Hunter I., Engel J. Structure and function of laminin: anatomy of a multidomain glycoprotein. FASEB J. 1990 Feb 1;4(2):148–160. doi: 10.1096/fasebj.4.2.2404817. [DOI] [PubMed] [Google Scholar]
- Bodey G. P. The emergence of fungi as major hospital pathogens. J Hosp Infect. 1988 Feb;11 (Suppl A):411–426. doi: 10.1016/0195-6701(88)90220-4. [DOI] [PubMed] [Google Scholar]
- Bodey G. P., Vartivarian S. Aspergillosis. Eur J Clin Microbiol Infect Dis. 1989 May;8(5):413–437. doi: 10.1007/BF01964057. [DOI] [PubMed] [Google Scholar]
- Bouchara J. P., Bouali A., Tronchin G., Robert R., Chabasse D., Senet J. M. Binding of fibrinogen to the pathogenic Aspergillus species. J Med Vet Mycol. 1988;26(6):327–334. [PubMed] [Google Scholar]
- Bouchara J. P., Oumeziane N. A., Lissitzky J. C., Larcher G., Tronchin G., Chabasse D. Attachment of spores of the human pathogenic fungus Rhizopus oryzae to extracellular matrix components. Eur J Cell Biol. 1996 May;70(1):76–83. [PubMed] [Google Scholar]
- Bouchara J. P., Tronchin G., Annaix V., Robert R., Senet J. M. Laminin receptors on Candida albicans germ tubes. Infect Immun. 1990 Jan;58(1):48–54. doi: 10.1128/iai.58.1.48-54.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bouchara J. P., Tronchin G., Larcher G., Chabasse D. The search for virulence determinants in Aspergillus fumigatus. Trends Microbiol. 1995 Aug;3(8):327–330. doi: 10.1016/s0966-842x(00)88965-9. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Burke C., Mayo K. H., Skubitz A. P., Furcht L. T. 1H NMR and CD secondary structure analysis of cell adhesion promoting peptide F-9 from laminin. J Biol Chem. 1991 Oct 15;266(29):19407–19412. [PubMed] [Google Scholar]
- CLARK H. F., SHEPARD C. C. A DIALYSIS TECHNIQUE FOR PREPARING FLUORESCENT ANTIBODY. Virology. 1963 Aug;20:642–644. doi: 10.1016/0042-6822(63)90292-7. [DOI] [PubMed] [Google Scholar]
- Casta e Silva Filho F., de Souza W., Lopes J. D. Presence of laminin-binding proteins in trichomonads and their role in adhesion. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8042–8046. doi: 10.1073/pnas.85.21.8042. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chammas R., Jasiulionis M. G., Jin F., Villa-Verde D. M., Reinhold V. N. Carbohydrate-binding proteins in cell-matrix interactions. Braz J Med Biol Res. 1994 Sep;27(9):2169–2179. [PubMed] [Google Scholar]
- Chen T., Belland R. J., Wilson J., Swanson J. Adherence of pilus- Opa+ gonococci to epithelial cells in vitro involves heparan sulfate. J Exp Med. 1995 Aug 1;182(2):511–517. doi: 10.1084/jem.182.2.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coulot P., Bouchara J. P., Renier G., Annaix V., Planchenault C., Tronchin G., Chabasse D. Specific interaction of Aspergillus fumigatus with fibrinogen and its role in cell adhesion. Infect Immun. 1994 Jun;62(6):2169–2177. doi: 10.1128/iai.62.6.2169-2177.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doolittle R. F. Fibrinogen and fibrin. Annu Rev Biochem. 1984;53:195–229. doi: 10.1146/annurev.bi.53.070184.001211. [DOI] [PubMed] [Google Scholar]
- Fraser D. W., Ward J. I., Ajello L., Plikaytis B. D. Aspergillosis and other systemic mycoses. The growing problem. JAMA. 1979 Oct 12;242(15):1631–1635. [PubMed] [Google Scholar]
- Furtado G. C., Slowik M., Kleinman H. K., Joiner K. A. Laminin enhances binding of Toxoplasma gondii tachyzoites to J774 murine macrophage cells. Infect Immun. 1992 Jun;60(6):2337–2342. doi: 10.1128/iai.60.6.2337-2342.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerson S. L., Talbot G. H., Hurwitz S., Strom B. L., Lusk E. J., Cassileth P. A. Prolonged granulocytopenia: the major risk factor for invasive pulmonary aspergillosis in patients with acute leukemia. Ann Intern Med. 1984 Mar;100(3):345–351. doi: 10.7326/0003-4819-100-3-345. [DOI] [PubMed] [Google Scholar]
- Ghosh A., Kole L., Bandyopadhyay K., Sarkar K., Das P. K. Evidence of a laminin binding protein on the surface of Leishmania donovani. Biochem Biophys Res Commun. 1996 Sep 4;226(1):101–106. doi: 10.1006/bbrc.1996.1317. [DOI] [PubMed] [Google Scholar]
- Gil M. L., Peñalver M. C., Lopez-Ribot J. L., O'Connor J. E., Martinez J. P. Binding of extracellular matrix proteins to Aspergillus fumigatus conidia. Infect Immun. 1996 Dec;64(12):5239–5247. doi: 10.1128/iai.64.12.5239-5247.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hinek A. Nature and the multiple functions of the 67-kD elastin-/laminin binding protein. Cell Adhes Commun. 1994 Jul;2(3):185–193. doi: 10.3109/15419069409004436. [DOI] [PubMed] [Google Scholar]
- Klapholz A., Salomon N., Perlman D. C., Talavera W. Aspergillosis in the acquired immunodeficiency syndrome. Chest. 1991 Dec;100(6):1614–1618. doi: 10.1378/chest.100.6.1614. [DOI] [PubMed] [Google Scholar]
- Klotz S. A. Adherence of Candida albicans to components of the subendothelial extracellular matrix. FEMS Microbiol Lett. 1990 Mar 15;56(3):249–254. doi: 10.1016/s0378-1097(05)80049-7. [DOI] [PubMed] [Google Scholar]
- Klotz S. A., Smith R. L. Glycosaminoglycans inhibit Candida albicans adherence to extracellular matrix proteins. FEMS Microbiol Lett. 1992 Dec 1;78(2-3):205–208. doi: 10.1016/0378-1097(92)90026-k. [DOI] [PubMed] [Google Scholar]
- Knibbs R. N., Perini F., Goldstein I. J. Structure of the major concanavalin A reactive oligosaccharides of the extracellular matrix component laminin. Biochemistry. 1989 Jul 25;28(15):6379–6392. doi: 10.1021/bi00441a034. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lissitzky J. C., Bouzon M., Loret E., Poupon M. F., Martin P. M. Laminin-mediated adhesion in metastatic rat rhabdomyosarcoma cell lines involves prominent interactions with the laminin E8 fragment. Clin Exp Metastasis. 1989 Jul-Aug;7(4):469–480. doi: 10.1007/BF01753667. [DOI] [PubMed] [Google Scholar]
- McMahon J. P., Wheat J., Sobel M. E., Pasula R., Downing J. F., Martin W. J., 2nd Murine laminin binds to Histoplasma capsulatum. A possible mechanism of dissemination. J Clin Invest. 1995 Aug;96(2):1010–1017. doi: 10.1172/JCI118086. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ollert M. W., Söhnchen R., Korting H. C., Ollert U., Bräutigam S., Bräutigam W. Mechanisms of adherence of Candida albicans to cultured human epidermal keratinocytes. Infect Immun. 1993 Nov;61(11):4560–4568. doi: 10.1128/iai.61.11.4560-4568.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Patti J. M., Allen B. L., McGavin M. J., Hök M. MSCRAMM-mediated adherence of microorganisms to host tissues. Annu Rev Microbiol. 1994;48:585–617. doi: 10.1146/annurev.mi.48.100194.003101. [DOI] [PubMed] [Google Scholar]
- Pendland J. C., Boucias D. G. Variations in the ability of galactose and mannose-specific lectins to bind to cell wall surfaces during growth of the insect pathogenic fungus Paecilomyces farinosus. Eur J Cell Biol. 1993 Apr;60(2):322–330. [PubMed] [Google Scholar]
- Peñalver M. C., O'Connor J. E., Martinez J. P., Gil M. L. Binding of human fibronectin to Aspergillus fumigatus conidia. Infect Immun. 1996 Apr;64(4):1146–1153. doi: 10.1128/iai.64.4.1146-1153.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plow E. F., Marguerie G. Inhibition of fibrinogen binding to human platelets by the tetrapeptide glycyl-L-prolyl-L-arginyl-L-proline. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3711–3715. doi: 10.1073/pnas.79.12.3711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Townsend R. R., Hilliker E., Li Y. T., Laine R. A., Bell W. R., Lee Y. C. Carbohydrate structure of human fibrinogen. Use of 300-MHz 1H-NMR to characterize glycosidase-treated glycopeptides. J Biol Chem. 1982 Aug 25;257(16):9704–9710. [PubMed] [Google Scholar]
- Tronchin G., Bouchara J. P., Larcher G., Lissitzky J. C., Chabasse D. Interaction between Aspergillus fumigatus and basement membrane laminin: binding and substrate degradation. Biol Cell. 1993;77(2):201–208. doi: 10.1016/s0248-4900(05)80189-3. [DOI] [PubMed] [Google Scholar]
- Tronchin G., Esnault K., Renier G., Filmon R., Chabasse D., Bouchara J. P. Expression and identification of a laminin-binding protein in Aspergillus fumigatus conidia. Infect Immun. 1997 Jan;65(1):9–15. doi: 10.1128/iai.65.1.9-15.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Valkonen K. H., Ringner M., Ljungh A., Wadström T. High-affinity binding of laminin by Helicobacter pylori: evidence for a lectin-like interaction. FEMS Immunol Med Microbiol. 1993 Jun;7(1):29–37. doi: 10.1111/j.1574-695X.1993.tb00378.x. [DOI] [PubMed] [Google Scholar]
- Vicentini A. P., Gesztesi J. L., Franco M. F., de Souza W., de Moraes J. Z., Travassos L. R., Lopes J. D. Binding of Paracoccidioides brasiliensis to laminin through surface glycoprotein gp43 leads to enhancement of fungal pathogenesis. Infect Immun. 1994 Apr;62(4):1465–1469. doi: 10.1128/iai.62.4.1465-1469.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Virkola R., Parkkinen J., Hacker J., Korhonen T. K. Sialyloligosaccharide chains of laminin as an extracellular matrix target for S fimbriae of Escherichia coli. Infect Immun. 1993 Oct;61(10):4480–4484. doi: 10.1128/iai.61.10.4480-4484.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vázquez-Juárez R., Ascencio F., Andlid T., Gustafsson L., Wadström T. The expression of potential colonization factors of yeasts isolated from fish during different growth conditions. Can J Microbiol. 1993 Dec;39(12):1135–1141. doi: 10.1139/m93-171. [DOI] [PubMed] [Google Scholar]
- Yamada K. M. Adhesive recognition sequences. J Biol Chem. 1991 Jul 15;266(20):12809–12812. [PubMed] [Google Scholar]
