This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wiker, H. G. Articles by Harboe, M. Search for Related Content PubMed PubMed Citation Articles by Wiker, H. G. Articles by Harboe, M.

 Previous Article  |  Next Article 

Infect Immun, April 1998, p. 1445-1452, Vol. 66, No. 4
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Immunochemical Characterization of the MPB70/80 and MPB83 Proteins of Mycobacterium bovis

Harald G. Wiker,^1,* Konstantin P. Lyashchenko,² A. Murat Aksoy,¹ Kenneth A. Lightbody,³ John M. Pollock,³ Sergiy V. Komissarenko,⁴ Sergiy O. Bobrovnik,⁴ Irina N. Kolesnikova,⁴ Leonid O. Mykhalsky,⁵ Maria L. Gennaro,² and Morten Harboe¹

Institute of Immunology and Rheumatology, University of Oslo, Oslo, Norway¹; Public Health Research Institute, New York²; Department of Agriculture for Northern Ireland, Veterinary Sciences Division, United Kingdom³; and Institute of Biochemistry⁴ and Kiev State University,⁵ Kiev, Ukraine

Received 29 September 1997/Returned for modification 10 November 1997/Accepted 8 January 1998

MPB70 and MPB80 (MPB70/80) and MPB83 are closely related antigens which are highly expressed in Mycobacterium bovis. MPB70/80 are soluble secreted antigens, while MPB83 is an exported lipoprotein associated with the bacterial surface. In the present study, these antigens had different mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. These differences may be explained by the fact that MPB70 and MPB83 both have two internal cysteine residues which would create ring structures by disulfide bonding. We analyzed the structures of MPB70/80 and MPB83 by using monoclonal antibodies (MAbs) raised against bovine purified protein derivative or whole M. bovis cells. MAb 1-5C reacted specifically with MPB70 and MPB80, and MAb MBS43 reacted specifically with MPB83, while the other antibodies, including several previously described MAbs, bound all three antigens. MAbs and polyclonal antibodies reacted strongly with reduced protein and less well with nonreduced protein, indicating involvement of linear epitopes. Epitopes of MAbs Bov-1, 2-6B, 1-5C, and 1-1D were mapped by using synthetic peptides of MPB70. Sequence comparison showed the peptide with the 1-5C-reactive epitope to have three residues different from those in the homologous region of MPB83. Exchanges of A for S in position 112 or Q for E in position 116 abolished the reactivity of MAb 1-5C. Polyclonal rabbit antibodies to native purified MPB70 reacted strongly with peptides 6, 7, and 8 of the N-terminal half of mature MPB70. Cattle sera of experimentally M. bovis-infected animals recognized a broader spectrum of peptides. These findings indicate that there is diagnostic potential for these proteins and that there is also a possible role for antibodies in elucidation of the host-mycobacterium relationship involving a surface-bound and exposed lipoprotein, MPB83, and its highly homologous soluble secreted MPB70/80 counterparts.

---

^* Corresponding author. Mailing address: Institute of Immunology and Rheumatology, University of Oslo, Fr. Qvamsgt. 1, N-0172 Oslo, Norway. Phone: 47 22 03 31 80. Fax: 47 22 20 72 87. E-mail: h.g.wiker{at}labmed.uio.no.

This article has been cited by other articles:

• Meikle, V., Alito, A., Llera, A. S., Gioffre, A., Peralta, A., Buddle, B. M., Cataldi, A. (2009). Identification of Novel Mycobacterium bovis Antigens by Dissection of Crude Protein Fractions. CVI 16: 1352-1359 [Abstract] [Full Text]  
• Dutton, R. J., Boyd, D., Berkmen, M., Beckwith, J. (2008). Bacterial species exhibit diversity in their mechanisms and capacity for protein disulfide bond formation. Proc. Natl. Acad. Sci. USA 105: 11933-11938 [Abstract] [Full Text]  
• Sweeney, F. P., Courtenay, O., Hibberd, V., Hewinson, R. G., Reilly, L. A., Gaze, W. H., Wellington, E. M. H. (2007). Environmental Monitoring of Mycobacterium bovis in Badger Feces and Badger Sett Soil by Real-Time PCR, as Confirmed by Immunofluorescence, Immunocapture, and Cultivation. Appl. Environ. Microbiol. 73: 7471-7473 [Abstract] [Full Text]  
• Pollock, J. M., McNair, J., Bassett, H., Cassidy, J. P., Costello, E., Aggerbeck, H., Rosenkrands, I., Andersen, P. (2003). Specific Delayed-Type Hypersensitivity Responses to ESAT-6 Identify Tuberculosis-Infected Cattle. J. Clin. Microbiol. 41: 1856-1860 [Abstract] [Full Text]  
• Al-Attiyah, R., Shaban, F. A., Wiker, H. G., Oftung, F., Mustafa, A. S. (2003). Synthetic Peptides Identify Promiscuous Human Th1 Cell Epitopes of the Secreted Mycobacterial Antigen MPB70. Infect. Immun. 71: 1953-1960 [Abstract] [Full Text]  
• Rhodes, S. G., Gavier-Widen, D., Buddle, B. M., Whelan, A. O., Singh, M., Hewinson, R. G., Vordermeier, H. M. (2000). Antigen Specificity in Experimental Bovine Tuberculosis. Infect. Immun. 68: 2573-2578 [Abstract] [Full Text]  
• Musser, J. M., Amin, A., Ramaswamy, S. (2000). Negligible Genetic Diversity of Mycobacterium tuberculosis Host Immune System Protein Targets: Evidence of Limited Selective Pressure. Genetics 155: 7-16 [Abstract] [Full Text]