title: First identification and functional characterization of an immunogenic protein in unculturable haemotrophic Mycoplasmas (Mycoplasma suis HspA1).
abstract: The antigenic structures of the haemotrophic Mycoplasma suis, an epicellular parasite of porcine erythrocytes, are largely unknown due to its unculturability. In this study, serological proteome and mass spectrometry analyses allowed the characterization of M. suis proteins targeted by the porcine antibody response: two proteins with characteristics of heat shock proteins, two proteins with characteristics of glycolytic enzymes, a RNA helicase- and an actin-like protein. The DnaK-like protein of M. suis (HspA1) was further analysed genetically and functionally. Its encoding gene (M. suis a1 gene) is 1.830 bp in size and corresponds to a 67 kDa protein. Immunoelectron microscopy verified the surface accessibility of HspA1 in M. suis. Recombinant HspA1 expressed in Escherichia coli demonstrated ATPase activity and antigenicity in experimentally infected pigs. In conclusion, this first identification and recombinant expression of an antigenic protein of M. suis provides the basis for the development of vaccines and new in vitro diagnostic assays.
authors.affiliation: Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland. lhoelzle@vetbakt.unizh.ch
authors.forename: Ludwig E
authors.initials: LE
authors.lastname: Hoelzle
authors.forename: Katharina
authors.initials: K
authors.lastname: Hoelzle
authors.forename: Alois
authors.initials: A
authors.lastname: Harder
authors.forename: Mathias
authors.initials: M
authors.lastname: Ritzmann
authors.forename: Heike
authors.initials: H
authors.lastname: Aupperle
authors.forename: Heinz-Adolf
authors.initials: HA
authors.lastname: Schoon
authors.forename: Karl
authors.initials: K
authors.lastname: Heinritzi
authors.forename: Max M
authors.initials: MM
authors.lastname: Wittenbrink
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009174:Mycoplasma / Q000235:genetics / Q000276:immunology*; D016366:Open Reading Frames; D020543:Proteome / Q000032:analysis / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; D020543:Proteome; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases