[en] A significant decrease in adherence rates of Mycoplasma bovis to bovine bronchial epithelial (BBE) cells has been observed after passage of the organism in artificial medium. Analysis of the proteins expressed by M. bovis isolate 2610 by two-dimensional (2-D) electrophoresis demonstrated differences between the cells harvested after the 7th and 116th passage. Three silver-stained prominent spots observed in 2-D electrophoretic separation of protein extracts of the lower-passaged cells were considerably less strongly expressed in the sample from higher-passaged cells. These spots had a molecular mass of approximately 24 kDa and an isoelectric point of about 5. The mass spectrometry analysis of these trypsin-sensitive proteins led to their identification as a unique new member of the Vsps family of membrane-associated proteins. Serum from a mouse immunized with these proteins significantly reduced adherence of M. bovis to BBE cells. This result underlines the function of this new Vsp in adherence of M. bovis to host cells. (c) 2005 Elsevier SAS. All rights reserved.
Disciplines :
Microbiology Veterinary medicine & animal health
Author, co-author :
Thomas, Anne; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Bactériologie et pathologie des maladies bactériennes
Leprince, Pierre ; Université de Liège - ULiège > CNCM/ Centre fac. de rech. en neurobiologie cell. et moléc.
Dizier, Isabelle ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Bactériologie et pathologie des maladies bactériennes
Gevaert, Kris; Flanders Interuniversity Institute of Biotechnology > Department of Medical Protein Research
Van Damme, Jozef; Flanders Interuniversity Institute of Biotechnology > Department of Medical Protein Research
Mainil, Jacques ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Bactériologie et pathologie des maladies bactériennes
Linden, Annick ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Santé et pathologies de la faune sauvage
Language :
English
Title :
Identification by two-dimensional electrophoresis of a new adhesin expressed by a low-passaged strain of Mycoplasma bovis
Publication date :
2005
Journal title :
Research in Microbiology
ISSN :
0923-2508
eISSN :
1769-7123
Publisher :
Elsevier, Paris, France
Volume :
156
Issue :
5-6,
Pages :
713-718
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
COST - European Cooperation in Science and Technology
Funders :
Ministère Fédéral de l'Agriculture F.R.S.-FNRS - Fonds de la Recherche Scientifique FWO - Fonds Wetenschappelijk Onderzoek Vlaanderen DG RDT - Commission Européenne. Direction Générale de la Recherche et de l'Innovation
H.J. Ball, D. Finlay, and G.A.C. Reilly Sandwich ELISA detection of Mycoplasma bovis in pneumonic calf lungs and nasal swabs Vet. Rec. 13 1994 531 532
T. Beier, H. Hotzel, I. Lysnyansky, C. Grajetzki, M. Heller, B. Rabeling, D. Yogev, and K. Sachse Intraspecies polymorphism of vsp genes and expression profiles of variable surface protein antigens (Vsps) in field isolates of Mycoplasma bovis Vet. Microbiol. 63 1998 189 203
N.C. Cary SAS System for Linear Models third ed. 1991 SAS Institute Inc. Cary, NC pp. 329
Y. Chavez Gonzalez, C. Ros Bascunana, G. Bolske, J.G. Mattson, C. Fernandez Molina, and K.-E. Johansson In vitro amplification of the 16SrRNA genes from Mycoplasma bovis and Mycoplasma agalactiae by PCR Vet. Microbiol. 47 1995 183 190
B. Fleury, D. Bergonier, X. Berthelot, E. Peterhans, J. Frey, and E. Vilei Characterization of P40, a cytoadhesin of Mycoplasma agalactiae Infect. Immun. 70 2002 5612 5621
A. Görg, G. Boguth, C. Obermaier, and W. Weiss Two-dimensional electrophoresis of proteins in an immobilized pH 4-12 gradient Electrophoresis 19 1998 516 1519
E.J. Hansen, R.M. Wilson, and J.B. Baseman Two-dimensional gel electrophoretic comparison of proteins from virulent and avirulent strains of Mycoplasma pneumoniae Infect. Immun. 24 1979 468 475
J. Heukeshoven, and R. Dernick Improved silver staining procedure for fast staining in PhastSystem Development Unit: I. Staining of sodium dodecyl sulfate gels Electrophoresis 9 1988 28 32
S.A. Leigh, and K.S. Wise Identification and functional mapping of Mycoplasma fermentans P29 adhesin Infect. Immun. 70 1999 4925 4935
L. Papazisi, K.E. Troy, T.S. Gorton, X. Liao, and S.J. Geary Analysis of cytoadherence-deficient, GapA-negative Mycoplasma gallisepticum strain R Infect. Immun. 68 2000 6643 6649
H. Pfützner, and K. Sachse Mycoplasma bovis as an agent of mastitis, pneumonia, arthritis and genital disorders in cattle Rev. Sci. Techn. OIE 15 1996 1477 1494
F. Poumarat, M. Perrin, and D. Longchambon Identification of ruminant mycoplasmas by dot immunobinding on membrane filtration (MF dot) Vet. Microbiol. 29 1991 329 338
S. Razin Adherence of pathogenic mycoplasmas to host cells Bioscience Rep. 19 1999 367 372
J. Regula, B. Ueberle, G. Boguth, A. Gorg, M. Schnolzer, R. Herrmann, and R. Frank Towards a two-dimensional proteome map of Mycoplasma pneumoniae Electrophoresis 21 2000 3765 3780
K. Sachse, H. Pfützner, M. Heller, and I. Hanel Inhibition of Mycoplasma bovis cytoadherence by a monoclonal antibody and various carbohydrate substances Vet. Microbiol. 36 1993 307 316
K. Sachse, C. Grajetzki, R. Rosengarten, I. Hanel, M. Heller, and H. Pfützner Mechanisms and factors involved in Mycoplasma bovis adhesion to host cells Zbl. Bakteriol. 284 1996 80 92
K. Sachse, J.H. Helbig, I. Lysnyansky, C. Grajetzki, W. Muller, E. Jacobs, and D. Yogev Epitope mapping of immunogenic and adhesive structures in repetitive domains of Mycoplasma bovis variable surface lipoproteins Infect. Immun. 68 2000 680 687
S. Subramaniam, D. Bergonier, F. Poumarat, S. Capaul, Y. Schlatter, J. Nicolet, and J. Frey Species identification of Mycoplasma bovis and Mycoplasma agalactiae based on the uvrC genes by PCR Mol. Cell. Probe 12 1998 161 169
A. Thomas, K. Sachse, I. Dizier, C. Grajetzki, F. Farnir, J.G. Mainil, and A. Linden Adherence to various host cell lines of Mycoplasma bovis strains differing in pathogenic and cultural features Vet. Microbiol. 91 2003 101 113
A. Thomas, K. Sachse, I. Dizier, H. Ball, J. Mainil, and A. Linden Adherence of Mycoplasma bovis strains to primary culture of bovine bronchial epithelial cells Microb. Pathogenesis 34 2003 141 148
C.J. Thorns, and E. Boughton Effect of serial passages through liquid medium on the virulence of Mycoplasma bovis for the mouse mammary gland Res. Vet. Sci. 29 1980 328 332
