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; Joachim Frey

doi:doi:10.1051/vetres/2009018

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Volume 40 / No 4 (July-August 2009)

Vet. Res., 40 4 (2009) 35

Abstract

Open Access

Issue		Vet. Res. Volume 40, Number 4, July-August 2009


Number of page(s)		12
DOI		https://doi.org/10.1051/vetres/2009018
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

Corresponding author: edy.vilei@vbi.unibe.ch

© INRA, EDP Sciences 2009