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Comparative proteomic analysis of pathogenic and non-pathogenic strains from the swine pathogen Mycoplasma
Paulo M Pinto1, Cátia S Klein, Arnaldo Zaha
1Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, UFRGS, Porto Alegre, RS, Brazil.
Background:
Mycoplasma 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.
Results:
In 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.
Conclusions:
Our 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.
Insights
Pathogenic Mycoplasma hyopneumoniae strains overexpress adhesins compared to non-pathogenic strains, enabling early host cell adhesion and infection. This proteomic difference explains their distinct lifestyles and infectivity in swine.
Area of Science:
- Veterinary Microbiology
- Bacterial Pathogenesis
- Proteomics
Background:
- Mycoplasma hyopneumoniae causes enzootic pneumonia (EP) in swine.
- Previous proteomic studies focused on pathogenic strain 7448.
- Comparative analysis of pathogenic and non-pathogenic strains is needed.
Purpose of the Study:
- To compare protein expression profiles of pathogenic and non-pathogenic Mycoplasma hyopneumoniae strains.
- To identify proteins associated with pathogenicity.
- To understand the physiological basis of M. hyopneumoniae infectivity.
Main Methods:
- Comparative proteomic analysis using 2DE and LC-MS/MS.
- Differential protein expression analysis between three M. hyopneumoniae strains (J, 7448, 7422).
- 2DE immunoblot analysis of P97 adhesin.
Main Results:
- 67 proteins showed differential expression between strains.
- Pathogenic strains overexpressed cytoadherence-related proteins, including P97 adhesin.
- LC-MS/MS identified 35% of the M. hyopneumoniae genome's coding capacity, with strain-specific and differentially abundant proteins.
Conclusions:
- Physiological differences exist between pathogenic and non-pathogenic M. hyopneumoniae strains.
- Pathogenic strains constitutively express adhesins for enhanced adhesion and infection.
- These proteomic differences explain the distinct lifestyles and infectivity of M. hyopneumoniae strains.
