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Doctoral Thesis
DOI
https://doi.org/10.11606/T.46.2003.tde-25092006-103444
Document
Author
Full name
Miryam Marroquin Quelopana
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2003
Supervisor
Committee
Alves, Maria Júlia Manso (President)
Miranda, Maria Teresa Machini de
Mortara, Renato Arruda
Silva, Ana Maria Moura da
Zanata, Sílvio Marques
Title in Portuguese
Trypanosoma cruzi e a interação com a matriz extracelular: modelagem da proteína Tc85-11 e determinação do sítio de ligação a laminina
Keywords in Portuguese
Laminina
Moleculas de adesão
Tc85-11
Trypanosoma cruzi
Abstract in Portuguese
Trypanosoma cruzi expressa um grupo de glicoproteínas de superfície, as Tc85, que pertencem à superfamília gênica das gp85/trans-sialidases. Neste trabalho mostramos um modelo de estrutura para um membro desta família, a proteína Tc85-11, a qual tem propriedades adesivas para laminina e para a superfície da célula. Esta estrutura consiste em um domínio N-terminal com pregamento ß-propeller e um domínio C-terminal ß-sandwich conectados por uma um α-hélice longa. A proteína recombinante que corresponde ao domínio amino terminal (Tc85-N), mas não ao domínio carboxi-terminal (Tc85-C), liga-se a laminina de uma maneira específica. Cinco peptídeos sintéticos, contidos no domínio N-terminal, aderem na superfície das células LLC-MK2 e inibem a infecção pelo T. cruzi. Dois destes peptídeos também podem inibir a interação Tc85-N - laminina de modo específico e poderiam representar o sítio de ligação a laminina. Estes resultados reforçam a hipótese que a Tc85-11 é uma proteína multi-adesiva, já que o domínio C-terminal, liga-se a citoqueratina-18. Por outro lado, o tratamento de células em cultura com Tc85-N aumenta a expressão de laminina, efeito já observado in vivo e quando culturas de células foram incubadas com antígenos liberados pelo parasita. A Tc85-11 pode ter, por tanto, um papel relevante na interação do parasita com o hospedeiro modulando ainda a expressão de componentes da matriz extracelular.
Title in English
Trypanosoma cruzi interaction with the extra-cellular matrix: modeling the TC85-11 protein and mapping the laminin-binding site
Keywords in English
Adhesionmolecules
Laminin
Tc85-11
Trypanosoma cruzi
Abstract in English
Trypanosoma cruzi expresses the Tc85 proteins, a set of surface glycoproteins belonging to the gp85/trans-sialidase supergene family. In this report we show a structure model for Tc85-11 a member of this family, which has adhesive properties to laminin and to the host cell surfaces. That structure consists in an N-terminus β-propeller and a C-terminus β-sandwich domains connected by a long α-helix. The recombinant protein corresponding to the N-domain (Tc85-N), but not to the C-domain (Tc85-C), was able to bind laminin in a specific manner. Five synthetic 20-mer peptides from the N-domain adhere onto LLC-MK2 cell surface and inhibit the T. cruzi infection. Two of these peptides can also inhibit specifically Tc85-N - laminin interaction and may represent the laminin-binding site. These results reinforce the hypothesis that the Tc85-11 protein is a multi-adhesive protein, since it also binds to citokeratin-18 by C-terminus domain. On the other hand, the treatment of host cells with Tc85-N increases the expression of laminin in the cell culture, as was previously reported for treatment with T. cruzi released antigens. In summary, Tc85-11 protein may play an important role in the host-parasite interaction, including the modulation of ECM expression.
 
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tese_Miryam_MQ.pdf (10.40 Mbytes)
Publishing Date
2008-05-05
 
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