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Doctoral Thesis
DOI
10.11606/T.87.2012.tde-05022013-084410
Document
Author
Full name
Ana Tung Ching Ching
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Ho, Paulo Lee (President)
Barbosa, Ângela Silva
Giordano, Ricardo José
Prudencio, Carlos Roberto
Ventura, Armando Morais
Title in Portuguese
Identificação de adesinas bacterianas por phage display.
Keywords in Portuguese
Bactérias (identificação)
Hamsters
Leptospirose
Ligantes
Proteínas recombinantes
Técnicas bacteriológicas
Abstract in Portuguese
A leptospirose é uma zoonose de importância mundial causada por bactérias do gênero Leptospira. No Brasil, a maioria dos casos é causada por L. interrogans sorovar Copenhageni. O objetivo destre trabalho foi identificar adesinas de leptospira pela técnica de Phage display. Bibliotecas com fragmentos genômicos resultaram na idendificação de ligantes de leptospira com afinidade por tecidos de hamster. Uma varredura dessas bibliotecas contra heparan sulfato proteoglicano (HSPG) identificou como ligantes as proteínas LigA e LigB. Proteínas recombinantes foram produzidas e submetidas à ligação às células de mamíferos e aos componentes de matriz extracelular. LigB recombinante foi capaz de se ligar ao HSPG, à heparina e às células de mamíferos. HSPG e heparina foram capazes de reduzir significativamente a interação dessa proteína com as células. Estes resultados evidenciam o papel de proteínas da leptospira na sua interação com o hospedeiro e ilustram a possibilidade do uso da técnica de phage display para identificar possíveis adesinas.
Title in English
Identification of bacterial adhesins through phage display.
Keywords in English
Bacteria (ID)
Bacteriological techniques
Binders
Hamsters
Leptospirosis
Recombinant proteins
Abstract in English
Leptospirosis is a worldwide important zoonosis caused by bacteria of the genus Leptospira. In Brazil, most cases is caused by L. interrogans serovar Copenhageni. Our goal was to identify leptospiras adhesins by phage display technique. Libraries of genomic fragments resulted in the identification of ligands with affinity for leptospiras hamster tissues. Screening these libraries against heparan sulfate proteoglycan (HSPG) identified the proteins LigA and LigB. Recombinant proteins were produced and subjected to binding to mammalian cells and extracellular matrix components. LigB recombinant was able to bind to HSPG, heparin and mammalian cells. HSPG and heparin were able to significantly reduce the interaction of this protein with cells. These results highlight the role of leptospiras proteins in its interaction with the host and illustrate the possibility of the use of phage display technique to identify potential adhesins.
 
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Publishing Date
2013-02-22
 
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