Identificação de proteínas de superfície de Staphylococcus saprophyticus e análise de fatores de virulência
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2014-06-09
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Universidade Federal de Goiás
Resumo
The Gram-positive bacterium Staphylococcus saprophyticus, one of the coagulasenegative
staphylococci, is the second most common causative agent of urinary tract
infection, affecting mainly sexually active women. Staphylococcus saprophyticus can
cause acute diseases as pyelonephritis, sepsis, nephrolithiasis, endocarditis,
urethritis, epididymitis and prostatitis. This work aims to identify Staphylococcus
saprophyticus surface proteins by using a proteolytic shaving approach, a
methodology that was established to identify surface-exposed protein domains by
tripsinization of intact cells. The peptides obtained were treated by trypsin, reduced,
alkylated and identified by nano-chromatography using a nanoACQUITY
UPLCTM system (Waters) coupled to a SYNAPT Q-TOF mass spectrometer (Waters).
The homology analysis was performed using the software ProteinLynx 2.3 (Waters).
Through the shaving, it was possible to identify 219 proteins, many of them,
described as virulence factors. Of total, 01 is cell wall protein, 09 are extracelular
proteins, 19 are membrane proteins and 190 are citoplasmatic proteins. Besides of
the lysis process, the presence of cytoplasmic proteins on cell surface can be due to
the activity of export pathways not yet identified and many of these proteins can be
proteins with moonlighting function, in other words, proteins that plays more of one
function, it can, in this case, plays functions on S. saprophyticus cell surface related
to bacterial virulence. The main proteins with moonlighting function include metabolic
enzymes of the glycolytic pathway; enzymes of other metabolic pathways, such as,
glyoxalate cycle; chaperones and proteins related with the proteic folding. The
prediction of cellular localization was performed through LocateP database. The
results of this research help to elucidate the strategies and machineries used by
proteins during the adhesion, infection and proliferation, leading us to understand the
interaction between the pathogenic bacteria S. saprophyticus and the human host.
The knowledge about the proteins present on the cell surface is of extreme
importance, because many of these proteins represent targets to new drugs,
therapeutic antibodies or vaccines, since the pathogen cell surface is the first to
contact with the host cells during the infection process.
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CARVALHO, A. J. Identificação de proteínas de superfície de Staphylococcus saprophyticus e análise de fatores de virulência. 2014. 66 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2014.