Comparative proteomic analysis of pathogenic and non-pathogenic strains from the swine pathogen Mycoplasma hyopneumoniae

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.

Characterization of Cleavage Events in the Multifunctional Cilium Adhesin Mhp684 (P146) Reveals a Mechanism by Which Mycoplasma hyopneumoniae Regulates Surface Topography

“…Most of the P97/P102 family members previously examined undergo at least one cleavage event and in some cases two or more cleavage events (19–22, 28, 29). The archetype cilium adhesin P97 itself undergoes extensive processing, creating many novel endoproteolytic fragments, but many of these fragments have not been experimentally characterized (20).…”

Recombinant Secreted Antigens from Mycoplasma hyopneumoniae Delivered as a Cocktail Vaccine Enhance the Immune Response of Mice

“…hyopneumoniae does not penetrate host cells, and thus it is believed that the pathogenesis of this microorganism is mediated by a complex and multifactorial process that involves components of the cell membrane and secreted proteins, many of which are still unidentified (5). Data generated from the sequencing of four isolates of M. hyopneumoniae (7448, 168, J, and 232) and the comparative genomic and proteomic analyses of pathogenic strains (7448 and 232) and a nonpathogenic strain (J) of M. hyopneumoniae (6,7,8,9) have allowed the identification of coding sequences (CDS) from secreted antigenic proteins and/or proteins involved in the pathogenicity of the microorganism. Some vaccines including only one of these candidate antigens have shown promising results (10,11).…”

Comparative proteomics of two Mycoplasma hyopneumoniae strains and Mycoplasma flocculare identified potential porcine enzootic pneumonia determinants

“…This suggests that these proteins are more fragmented in the cell surface than in in the cytoplasm, when they are expected to be mostly unprocessed. Previous studies have showed that adhesins are targets of post-translational proteolytic events [20–24] which can be differential between M. hyopneumoniae strains [15]. Along with differential adhesin abundance, the possibly differential adhesin post-translational proteolytic processing likely impact on bacterial pathogenicity and deserve further investigation.…”

“…However, previous comparative transcriptomic studies between M. hyopneumoniae and M. flocculare [14] failed to find differences in the relative transcription levels for most genes. On the other hand, pioneer proteomic studies, have provided evidences of differential protein abundance and post-translational processing between M. hyopneumoniae pathogenic (7448 and 7422) and non-pathogenic (J) strains [15]. Moreover, a recent comparative proteomics study between M. hyopneumoniae and M. flocculare secreted proteins revealed several virulence-related differences between these mycoplasma species [16].…”