Proteomic survey of the pathogenic Mycoplasma hyopneumoniae strain 7448 and identification of novel post-translationally modified and antigenic proteins.
Mycoplasma hyopneumoniae is an important pathogen for pigs, being the causative agent of enzootic pneumonia. Recently, the genome sequences of three strains, J, 7448 and 232 have been reported. Here, we describe the results of a proteomic analysis, based on two-dimensional gel electrophoresis of soluble protein extracts, immunoblot and mass spectrometry, which was carried out aiming the identification of gene products and antigenic proteins from the M. hyopneumoniae pathogenic strain 7448. A preliminary M. hyopneumoniae proteome map in two pH ranges (3-10 and 4-7) was produced. A total of 31 different coding DNA sequences (CDSs), including three hypothetical ones, were experimentally verified with the identification of the corresponding protein products by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. According to the Clusters of Orthologous Groups (COG) functional classification, the identified proteins were assigned to the groups of metabolism (13), cellular processes (5) and information and storage processing (4). Nine of the identified proteins were not classifiable by COG, including some related to cytoadherence and possibly involved in pathogenicity. Moreover, at least five highly antigenic proteins of M. hyopneumoniae were identified by immunoblots, including four novel ones (a heat shock protein 70, an elongation factor Tu, a pyruvate dehydrogenase E1-beta subunit and the P76 membrane protein). The now available proteome map is expected to serve as a reference for comparative analyses between M. hyopneumoniae pathogenic and non-pathogenic strains, and for methabolic studies based on cells cultured under modified conditions.
Pinto
Paulo Marcos
PM
Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, UFRGS, Porto Alegre, RS, Brazil.
Chemale
Gustavo
G
de Castro
Luiza Amaral
LA
Costa
Ana Paula Metz
AP
Kich
Jalusa Deon
JD
Vainstein
Marilene Henning
MH
Zaha
Arnaldo
A
Ferreira
Henrique Bunselmeyer
HB
D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000662:veterinary; D015151:Immunoblotting / Q000662:veterinary; D009175:Mycoplasma Infections / Q000382:microbiology / Q000662:veterinary*; D045705:Mycoplasma hyopneumoniae / Q000378:metabolism*; D045729:Pneumonia of Swine, Mycoplasmal / Q000382:microbiology*; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000662:veterinary
D001426:Bacterial Proteins