Mycobacterium tuberculosis, the causative agent of tuberculosis, produces a heparin-binding haemagglutinin adhesin (HBHA), which is involved in its epithelial adherence. To ascertain whether HBHA is also present in fast-growing mycobacteria, Mycobacterium smegmatis was studied using anti-HBHA monoclonal antibodies (mAbs). A cross-reactive protein was detected by immunoblotting of M. smegmatis whole-cell lysates. However, the M. tuberculosis HBHA-encoding gene failed to hybridize with M. smegmatis chromosomal DNA in Southern blot analyses. The M. smegmatis protein recognized by the anti-HBHA mAbs was purified by heparin-Sepharose chromatography, and its amino-terminal sequence was found to be identical to that of the previously described histone-like protein, indicating that M. smegmatis does not produce HBHA. Biochemical analysis of the M. smegmatis histone-like protein shows that it is glycosylated like HBHA. Immunoelectron microscopy demonstrated that the M. smegmatis protein is present on the mycobacterial surface, a cellular localization inconsistent with a histone-like function, but compatible with an adhesin activity. In vitro protein interaction assays showed that this glycoprotein binds to laminin, a major component of basement membranes. Therefore, the protein was called M. smegmatis laminin-binding protein (MS-LBP). MS-LBP does not appear to be involved in adherence in the absence of laminin but is responsible for the laminin-mediated mycobacterial adherence to human pneumocytes and macrophages. Homologous laminin-binding adhesins are also produced by virulent mycobacteria such as M. tuberculosis and Mycobacterium leprae, suggesting that this adherence mechanism may contribute to the pathogenesis of mycobacterial diseases.
Raviglione M. C., Global epidemiology of tuberculosis. Morbidity and mortality of a worldwide epidemic, 10.1001/jama.273.3.220
Noordeen S.K., Bull WHO, 70, 7 (1992)
Inderlied C B, Kemper C A, Bermudez L E, The Mycobacterium avium complex., 10.1128/cmr.6.3.266
Schlesinger L.S., J Immunol, 144, 2771 (1990)
Schlesinger L.S., J Immunol, 150, 2920 (1993)
Stokes R.W., J Immunol, 151, 7067 (1993)
Shepard C. C., Phagocytosis by HeLa Cells and Their Susceptibility to Infection by Human Tubercle Bacilli., 10.3181/00379727-90-22043
Mapother M.E., Infect Immun, 45, 67 (1984)
Bermudez L.E., Infect Immun, 64, 1400 (1996)
Menozzi F.D., Infect Immun, 62, 769 (1994)
Leong J.M., Infect Immun, 63, 874 (1995)
Compton Teresa, Nowlin Dawn M., Cooper Neil R., Initiation of Human Cytomegalovirus Infection Requires Initial Interaction with Cell Surface Heparan Sulfate, 10.1006/viro.1993.1192
Shieh M.-T., J Virol, 68, 1224 (1994)
Gysin Jürg, Pouvelle Bruno, Le Tonquèze Michèle, Edelman Lena, Boffa Marie-Claire, Chondroitin sulfate of thrombomodulin is an adhesion receptor for Plasmodium falciparum-infected erythrocytes, 10.1016/s0166-6851(97)00082-0
Ortega-Barria Eduardo, Boothroyd John C., AToxoplasmaLectin-like Activity Specific for Sulfated Polysaccharides Is Involved in Host Cell Infection, 10.1074/jbc.274.3.1267
Menozzi F. D., Identification of a heparin-binding hemagglutinin present in mycobacteria, 10.1084/jem.184.3.993
Menozzi F. D., Bischoff R., Fort E., Brennan M. J., Locht C., Molecular characterization of the mycobacterial heparin-binding hemagglutinin, a mycobacterial adhesin, 10.1073/pnas.95.21.12625
Delogu G., J Bacteriol, 181, 7464 (1999)
Pethe Kevin, Aumercier Marc, Fort Emmanuelle, Gatot Christophe, Locht Camille, Menozzi Franco D., Characterization of the Heparin-binding Site of the Mycobacterial Heparin-binding Hemagglutinin Adhesin, 10.1074/jbc.275.19.14273
Bermudez L.E., Shelton K., Young L.S., Comparison of the ability of Mycobacterium avium, M. smegmatis and M. tuberculosis to invade and replicate within HEp-2 epithelial cells, 10.1016/s0962-8479(05)80012-7
Timpl R., Engel J., Martin G.R., Laminin — a multifunctional protein of basement membranes, 10.1016/0968-0004(83)90213-x
Prabhakar S., Annapurna P.S., Annapurna P.S., Jain N.K., Dey A.B., Tyagi J.S., Prasad H.K., Identification of an immunogenic histone-like protein (HLPMt) ofMycobacterium tuberculosis, 10.1054/tuld.1998.0004
Shimoji Y., Ng V., Matsumura K., Fischetti V. A., Rambukkana A., A 21-kDa surface protein of Mycobacterium leprae binds peripheral nerve laminin-2 and mediates Schwann cell invasion, 10.1073/pnas.96.17.9857
Lee B. H., Murugasu-Oei B., Dick T., Upregulation of a histone-like protein in dormant Mycobacterium smegmatis, 10.1007/s004380050919
Nair J., Infect Immun, 61, 1074 (1993)
Yamada Hiroki, Denzer Alain J., Hori Hisae, Tanaka Takeshi, Anderson Louise V. B., Fujita Sachiko, Fukuta-Ohi Hiroko, Shimizu Teruo, Ruegg Markus A., Matsumura Kiichiro, Dystroglycan Is a Dual Receptor for Agrin and Laminin-2 in Schwann Cell Membrane, 10.1074/jbc.271.38.23418
Hoffman Matthew P., Nomizu Motoyoshi, Roque Eva, Lee Sharon, Jung Dale W., Yamada Yoshi, Kleinman Hynda K., Laminin-1 and Laminin-2 G-domain Synthetic Peptides Bind Syndecan-1 and Are Involved in Acinar Formation of a Human Submandibular Gland Cell Line, 10.1074/jbc.273.44.28633
Talts Jan F., Andac Zeynep, Göhring Walter, Brancaccio Andrea, Timpl Rupert, Binding of the G domains of laminin α1 and α2 chains and perlecan to heparin, sulfatides, α-dystroglycan and several extracellular matrix proteins, 10.1093/emboj/18.4.863
Sambrook J., Molecular Cloning: A Laboratory Manual (1989)
Rouse D.A., Infect Immun, 59, 2595 (1991)
Leininger E., J Infect Dis, 175, 1423 (1997)
Menozzi F.D., Infect Immun, 59, 3982 (1991)
Sweeley C. C., Bentley Ronald, Makita M., Wells W. W., , 10.1021/ja00899a032
LAEMMLI U. K., Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4, 10.1038/227680a0
Towbin H., Staehelin T., Gordon J., Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications., 10.1073/pnas.76.9.4350
Bradford M, A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, 10.1006/abio.1976.9999