Open Access
Vet. Res.
Volume 40, Number 4, July-August 2009
Number of page(s) 12
Published online 15 April 2009
How to cite this article Vet. Res. (2009) 40:35
How to cite this article: Vet. Res. (2009) 40:35
DOI: 10.1051/vetres/2009018

Functional and antigenic properties of GlpO from Mycoplasma mycoides subsp. mycoides SC: characterization of a flavin adenine dinucleotide-binding site deletion mutant

Daniela F. Bischof, Edy M. Vilei and Joachim Frey

Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Länggass-Strasse 122, P.O. Box, CH-3001 Bern, Switzerland

Received 18 February 2009; accepted 10 April 2009; published online 15 April 2009

Abstract - L-$\alpha$-glycerophosphate oxidase (GlpO) plays a central role in virulence of Mycoplasma mycoides subsp. mycoides SC, a severe bacterial pathogen causing contagious bovine pleuropneumonia (CBPP). It is involved in production and translocation of toxic H2O2 into the host cell, causing inflammation and cell death. The binding site on GlpO for the cofactor flavin adenine dinucleotide (FAD) has been identified as $\underline{\rm Gly}_{12}$–Gly13 $\underline{\rm Gly}_{14}$–Ile15–Ile16 $\underline{\rm Gly}_{17}$. Recombinant GlpO lacking these six amino acids (GlpO$\Delta$FAD) was unable to bind FAD and was also devoid of glycerophosphate oxidase activity, in contrast to non-modified recombinant GlpO that binds FAD and is enzymatically active. Polyclonal monospecific antibodies directed against GlpO$\Delta$FAD, similarly to anti-GlpO antibodies, neutralised H2O2 production of M. mycoides subsp. mycoides SC grown in the presence of glycerol, as well as cytotoxicity towards embryonic calf nasal epithelial (ECaNEp) cells. The FAD-binding site of GlpO is therefore suggested as a valuable target site for the future construction of deletion mutants to yield attenuated live vaccines of M. mycoides subsp. mycoides SC necessary to efficiently combat CBPP.

Key words: CBPP / L-$\alpha$-glycerophosphate oxidase / FAD-binding site / targeted deletion mutant / neutralizing Antibody

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© INRA, EDP Sciences 2009