Perfil comparativo do exoproteoma de 3 isolados clínicos de Staphylococcus saprophyticus
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2016-09-30
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Universidade Federal de Goiás
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Staphylococcus saprophyticus is a Gram-positive bacterium responsible for genitourinary infections, mainly affecting young sexually active women. Along with Escherichia coli is responsible for 90% of infections in fertile women, however, can cause infections in men and women of all ages. The repertoire of proteins secreted by pathogenic microorganisms is used to ensure success in the establishment of infection and persistence in the host. In this sense, this study aimed to comparative characterization of extracellular proteome of 3 clinical strain of S. saprophyticus, in order to detect possible differences in the secretion of proteins related to virulence and adaptation of the microorganism. The strains used in the study are called ATCC 15305, 7108 and 9325. Ultra-Performance Liquid Chromatography coupled to Mass Spectrometry in tandem (UPLC-MSE) have identified a total of 159 proteins. Among them, 44 were found in exoproteome of 3 strain, while 20 only in strains ATCC 15305 and 7108, 11 in ATCC 15305 and 9325, and 12 in 7108 and 9325. Fifteen peptides were expressed exclusively by S. saprophyticus 9325, 21 by ATCC 15305 strain and 36 by 7108. The three strain secreted molecules with biological function related to the glycolytic pathway, such as the triosephosphate isomerase (TPI), enolase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and fructose-bisphosphate aldolase, molecules clearly involved in extracellular processes from other pathogenic organisms, proteins known as "moonlighting". Proteins involved in defense against stress, such as catalase, alkyl hydroperoxide reductase subunit C (AhpC), superoxide dismutase (SOD), were also detected in the analysis, among others related to metabolic processes such as lactic fermentation, cell wall synthesis, protein processing, and iron metabolism. Staphylococcal secretory antigen A (SsaA), a immunogenic molecule previously detected in ATCC 15305 strain, was not detected in strain 7108, as confirmed by Western-blotting assay. PCR reactions with specific primers and genomic DNA of the 3 strains and subsequent sequencing ssaA gene, showed that the strain has the gene under conditions identical to those found in S. saprophyticus ATCC 15305 and 9325, but is not able to secrete the antigen into the extracellular milieu, under environmental conditions in study. In exoproteome S. saprophyticus 7108 was found to take a greater amount of unique proteins, including four of the class of peptidases, enzymes often identified as potential virulence factors of bacterial. Thus, the differences found in repertoire of extracellular proteins of three clinical strain of S. saprophyticus, demonstrate a metabolic flexibility used by this uropathogen to promote pathogenesis in human genitourinary tract.
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OLIVEIRA, A. S. Perfil comparativo do exoproteoma de 3 isolados clínicos de Staphylococcus saprophyticus. 2016. 100 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2016.