Identification by two-dimensional electrophoresis of a new adhesin expressed by a low-passaged strain of Mycoplasma bovis.
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.
Thomas
Anne
A
Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, B43A, Sart Tilman, 4000 Liège, Belgium. athomas@ulg.ac.be
Leprince
Pierre
P
Dizier
Isabelle
I
Ball
Hywel
H
Gevaert
Kris
K
Van Damme
Jozef
J
Mainil
Jacques
J
Linden
Annick
A
D018829:Adhesins, Bacterial / Q000096:biosynthesis* / Q000302:isolation & purification*; D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000302:isolation & purification; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D002417:Cattle; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000382:microbiology; D007526:Isoelectric Point; D013058:Mass Spectrometry; D008970:Molecular Weight; D045682:Mycoplasma bovis / Q000502:physiology*; D020543:Proteome / Q000032:analysis* / Q000302:isolation & purification
D018829:Adhesins, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D020543:Proteome; C121437:Vsp protein, bacterial