HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Pethe, K. Articles by Menozzi, F. D. Search for Related Content PubMed PubMed Citation Articles by Pethe, K. Articles by Menozzi, F. D. Social Bookmarking                      What's this?

J Biol Chem, Vol. 275, Issue 19, 14273-14280, May 12, 2000

Characterization of the Heparin-binding Site of the Mycobacterial Heparin-binding Hemagglutinin Adhesin^*

Kevin Pethe^§, Marc Aumercier^¶, Emmanuelle Fort, Christophe Gatot, Camille Locht^**, and Franco D. Menozzi

From the  INSERM U447, Mécanismes Moléculaires de la Pathogénie Microbienne, Institut Pasteur de Lille, Institut de Biologie de Lille, 1 rue A. Calmette, 59019 Lille Cedex, France, ^¶ CNRS, Mécanismes du Développement et de la Cancérisation, UMR 8526, Institut de Biologie de Lille, 59019 Lille Cedex, France, and the  Computer Cell Culture Center s.a., 14 rue de la Marlette, 7140 Seneffe, Belgium

The mycobacterial adhesin heparin-binding hemagglutinin (HBHA) contains several lysine-rich repeats at its carboxyl-terminal end. Using truncated recombinant HBHA forms and hybrid proteins containing HBHA repeats grafted onto the Escherichia coli maltose-binding protein (MBP), we found that these repeats are responsible for heparin binding. Immunofluorescence microscopy studies revealed that their deletion abrogates binding of HBHA to human pneumocytes. Conversely, when fused to MBP, the HBHA repeats confer pneumocyte adherence properties to the hybrid protein. Treatment of pneumocytes with glycosaminoglycan-degrading enzymes showed that HBHA binding depends on the presence of heparan sulfate chains on the cell surface. The epitope of a monoclonal antibody that inhibits mycobacterial adherence to epithelial cells was mapped within the lysine-rich repeats, confirming their involvement in mycobacterial adherence to epithelial cells. Surface plasmon resonance analyses showed that recombinant HBHA binds to immobilized heparin with fast association kinetics (k_a = 5.62 (± 0.10) × 10⁵ M¹ s¹), whereas the dissociation kinetics were slower (k_d = 0.015 (± 0.002) s¹), yielding a K_D value of 26 nM. Similar analyses with grafted MBP indicated similar kinetic constants, indicating that the carboxyl-terminal repeats contain the entire heparin-binding site of HBHA. The molecular characterization of the interactions of HBHA with epithelial glycosaminoglycans should help to better understand mycobacterial adherence within the lungs and may ultimately lead to new approaches for therapy or immunoprophylaxis.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				C. Verbelen, V. Dupres, D. Raze, C. Bompard, C. Locht, and Y. F. Dufrene Interaction of the Mycobacterial Heparin-Binding Hemagglutinin with Actin, as Evidenced by Single-Molecule Force Spectroscopy J. Bacteriol., December 1, 2008; 190(23): 7614 - 7620. [Abstract] [Full Text] [PDF]

				C. Esposito, M. V. Pethoukov, D. I. Svergun, A. Ruggiero, C. Pedone, E. Pedone, and R. Berisio Evidence for an Elongated Dimeric Structure of Heparin-Binding Hemagglutinin from Mycobacterium tuberculosis J. Bacteriol., July 1, 2008; 190(13): 4749 - 4753. [Abstract] [Full Text] [PDF]

				C. Verbelen, D. Raze, F. Dewitte, C. Locht, and Y. F. Dufrene Single-Molecule Force Spectroscopy of Mycobacterial Adhesin-Adhesin Interactions J. Bacteriol., December 15, 2007; 189(24): 8801 - 8806. [Abstract] [Full Text] [PDF]

				C. Jenkins, S. J. Geary, M. Gladd, and S. P. Djordjevic The Mycoplasma gallisepticum OsmC-like protein MG1142 resides on the cell surface and binds heparin Microbiology, May 1, 2007; 153(5): 1455 - 1463. [Abstract] [Full Text] [PDF]

				A-R. Shin, K.-S. Lee, J.-S. Lee, S.-Y. Kim, C.-H. Song, S.-B. Jung, C.-S. Yang, E.-K. Jo, J.-K. Park, T.-H. Paik, et al. Mycobacterium tuberculosis HBHA Protein Reacts Strongly with the Serum Immunoglobulin M of Tuberculosis Patients. Clin. Vaccine Immunol., August 1, 2006; 13(8): 869 - 875. [Abstract] [Full Text] [PDF]

				C. Jenkins, J. L. Wilton, F. C. Minion, L. Falconer, M. J. Walker, and S. P. Djordjevic Two Domains within the Mycoplasma hyopneumoniae Cilium Adhesin Bind Heparin Infect. Immun., January 1, 2006; 74(1): 481 - 487. [Abstract] [Full Text] [PDF]

				J. R. Hayman, T. R. Southern, and T. E. Nash Role of Sulfated Glycans in Adherence of the Microsporidian Encephalitozoon intestinalis to Host Cells In Vitro Infect. Immun., February 1, 2005; 73(2): 841 - 848. [Abstract] [Full Text] [PDF]

				I. R. Henderson, F. Navarro-Garcia, M. Desvaux, R. C. Fernandez, and D. Ala'Aldeen Type V Protein Secretion Pathway: the Autotransporter Story Microbiol. Mol. Biol. Rev., December 1, 2004; 68(4): 692 - 744. [Abstract] [Full Text] [PDF]

				K. Aoki, S. Matsumoto, Y. Hirayama, T. Wada, Y. Ozeki, M. Niki, P. Domenech, K. Umemori, S. Yamamoto, A. Mineda, et al. Extracellular Mycobacterial DNA-binding Protein 1 Participates in Mycobacterium-Lung Epithelial Cell Interaction through Hyaluronic Acid J. Biol. Chem., September 17, 2004; 279(38): 39798 - 39806. [Abstract] [Full Text] [PDF]

				J. P. Bannantine, J. F. J. Huntley, E. Miltner, J. R. Stabel, and L. E. Bermudez The Mycobacterium avium subsp. paratuberculosis 35 kDa protein plays a role in invasion of bovine epithelial cells Microbiology, August 1, 2003; 149(8): 2061 - 2069. [Abstract] [Full Text] [PDF]

				S. L. Mueller-Ortiz, E. Sepulveda, M. R. Olsen, C. Jagannath, A. R. Wanger, and S. J. Norris Decreased Infectivity despite Unaltered C3 Binding by a {Delta}hbhA Mutant of Mycobacterium tuberculosis Infect. Immun., December 1, 2002; 70(12): 6751 - 6760. [Abstract] [Full Text] [PDF]

				K. Pethe, P. Bifani, H. Drobecq, C. Sergheraert, A.-S. Debrie, C. Locht, and F. D. Menozzi Mycobacterial heparin-binding hemagglutinin and laminin-binding protein share antigenic methyllysines that confer resistance to proteolysis PNAS, August 6, 2002; 99(16): 10759 - 10764. [Abstract] [Full Text] [PDF]

				S. L. Mueller-Ortiz, A. R. Wanger, and S. J. Norris Mycobacterial Protein HbhA Binds Human Complement Component C3 Infect. Immun., December 1, 2001; 69(12): 7501 - 7511. [Abstract] [Full Text] [PDF]

				K. K. Singh, X. Zhang, A. S. Patibandla, P. Chien Jr., and S. Laal Antigens of Mycobacterium tuberculosis Expressed during Preclinical Tuberculosis: Serological Immunodominance of Proteins with Repetitive Amino Acid Sequences Infect. Immun., June 1, 2001; 69(6): 4185 - 4191. [Abstract] [Full Text] [PDF]