Abstract
BackgroundMycoplasma hyopneumoniae is a highly infectious swine pathogen and is the causative agent of enzootic pneumonia (EP). Following the previous report of a proteomic survey of the pathogenic 7448 strain of swine pathogen, Mycoplasma hyopneumoniae, we performed comparative protein profiling of three M. hyopneumoniae strains, namely the non-pathogenic J strain and the two pathogenic strains 7448 and 7422.ResultsIn 2DE comparisons, we were able to identify differences in expression levels for 67 proteins, including the overexpression of some cytoadherence-related proteins only in the pathogenic strains. 2DE immunoblot analyses allowed the identification of differential proteolytic cleavage patterns of the P97 adhesin in the three strains. For more comprehensive protein profiling, an LC-MS/MS strategy was used. Overall, 35% of the M. hyopneumoniae genome coding capacity was covered. Partially overlapping profiles of identified proteins were observed in the strains with 81 proteins identified only in one strain and 54 proteins identified in two strains. Abundance analysis of proteins detected in more than one strain demonstrates the relative overexpression of 64 proteins, including the P97 adhesin in the pathogenic strains.ConclusionsOur results indicate the physiological differences between the non-pathogenic strain, with its non-infective proliferate lifestyle, and the pathogenic strains, with its constitutive expression of adhesins, which would render the bacterium competent for adhesion and infection prior to host contact.
Highlights
Mycoplasma hyopneumoniae is a highly infectious swine pathogen and is the causative agent of enzootic pneumonia (EP)
Comparative shotgun proteomics of M. hyopneumoniae Since 2DE does not favour the identification of less abundant protein species or proteins with extremes of pI, we performed a complementary shotgun liquid chromatography (LC)-MS/MS analysis for a more comprehensive comparative analysis of the
Protein extracts from in vitro cultured strains were individually digested with trypsin and each of the resulting peptide mixtures were independently analysed three times by LC-MS/MS
Summary
Mycoplasma hyopneumoniae is a highly infectious swine pathogen and is the causative agent of enzootic pneumonia (EP). Mycoplasma hyopneumoniae is a highly infectious swine pathogen and is the causative agent of enzootic pneumonia (EP), a disease characterised by a sporadic, dry, and non-productive cough, retarded growth, and inefficient food conversion [1]. The M. hyopneumoniae genomes of a non-pathogenic (J) and two pathogenic strains (strains 7448 and 232) [2,3] have been sequenced. The mechanisms of M. hyopneumoniae pathogenicity suggest the existence of several other classes of unidentified virulence factors, such as genes/proteins involved in secretion and/or trafficking of molecules between host and pathogen cells, or evasion and/or modulation of the host immune system [6]. The observed presence of a putative integrative conjugal element (ICEH) in three M. hyopneumoniae pathogenic strains, but not in a non-pathogenic strain, suggests the involvement of potentially mobile genetic elements in the M. hyopneumoniae virulence [6,9]
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