Biochemical and antigenic characterisation of Mycoplasma gallisepticum membrane proteins P52 and P67 (pMGA).
Two membrane proteins from the avian pathogen Mycoplasma gallisepticum have been previously purified using a simple, efficient and non-denaturing method: a lipoprotein P67 (pMGA) and P52. In the current study, the lipid part of P67 was chemically analysed. The molecular structure of the lipoprotein-lipid component was determined to be S-glyceryl cysteine with two O-ester-linked acyl chains. Fatty acid analysis of the purified P67 indicated a heterogeneous composition: palmitic acid (16:0)>stearic acid (18:0)>oleic acid (18:1c)>myristic acid (14:0), with 16:0 as the major component. These findings, along with previously published results, support the conclusion that P67 is pMGA1.2, a true membrane-associated lipoprotein although not N-acylated. In contrast to P67, P52 is not a lipoprotein. Topological experiments using in situ treatment with proteases and growth inhibition in the presence of anti-P52 serum provided evidence of the surface exposition of the polypeptide. The N-terminal sequence of P52 was found to be similar to the dihydrolipoamide acetyltransferase from several mollicutes; this enzyme is a membrane-associated component of the pyruvate dehydrogenase complex. Immunoblotting techniques revealed that the surface antigens P52 and P67 were specific to the species M. gallisepticum and the closely related species M. imitans. No antigenic difference was revealed within these species with the anti-P52 serum, while anti-P67 serum confirmed the antigenic variability of P67. The potential of P52 and P67 as antigens in serological diagnosis tests or as candidates for anti-mycoplasma subunit vaccines is discussed.
Jan
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Université de Rennes I, UMR CNRS 6026 Groupe "Membranes et Osmorégulation", Avenue du Général Leclerc, 35042 Rennes Cedex, France.
Le Hénaff
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Fontenelle
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Wróblewski
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D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D016882:ISCOMs; D008074:Lipoproteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D016882:ISCOMs; D008074:Lipoproteins; D008565:Membrane Proteins; C114986:p67 protein, Mycoplasma