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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2012.tde-23042013-095557
Document
Author
Full name
Mauricio Scavassini Penã
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Carboni, Beatriz Simonsen Stolf (President)
Cotrim, Paulo Cesar
Uliana, Silvia Reni Bortolin
Title in Portuguese
Identificação de ligantes da metacaspase de Leishmania (Leishmania) amazonensis pela técnica de "Phage Display".
Keywords in Portuguese
Leishmania
Amastigotes
Apoptose
Proteínas do choque térmico
Protozoa
Abstract in Portuguese
Durante o ciclo de vida da Leishmania, amastigotas vivem no interior de fagolisossomas de células fagocíticas de hospedeiros vertebrados, enquanto promastigotas vivem no interior do vetor invertebrado. Proteases intracelulares como as caspases são as principais efetoras no processo apoptótico. Metacaspases (MCAs) são formas evolutivas distantes das caspases de metazoários, presentes em protozoários, plantas e fungos, e vistas como potenciais alvos para combate dos parasitas sem prejuízo do hospedeiro. Ligantes e moduladores das metacaspases são até hoje desconhecidos. O Phage Display é uma técnica baseada na expressão de proteínas sintéticas nos capsidíos de fagos, usada com o propósito de selecionar ligantes de proteínas, células ou tecidos. Produzimos a metacaspase recombinante de Leishmania L. amazonensis e aplicamos Phage Display para buscar peptídeos ligantes dessa enzima. Esses peptídeos permitiram identificar potenciais proteínas ligantes da MCA, como quinases e cinesinas, que fornecem informações sobre a regulação e controle de sua atividade. Futuramente testaremos se peptídeos ativadores da MCA poderão induzir apoptose do parasita e serem usados como drogas para o tratamento da leishmaniose.
Title in English
Identification of ligands of Leishmania (Leishmania) amazonensis metacaspase using Phage Display.
Keywords in English
Leishmania
Amastigote
Apoptosis
Heat shock proteins
Protozoa
Abstract in English
During its life cycle, Leishmania amastigotes live inside phagolysosomes of phagocytic cells of vertebrate hosts, while promastigotes live inside the invertebrate vector. Intracellular proteases such as caspases are key effectors in the apoptotic process. Metacaspases (MCAs) are distant evolutionary forms of metazoan caspases found in protozoa, plants and fungi, and seen as potential targets to destroy the parasites without damage to the host. Ligands and modulators of metacaspases are so far unknown. Phage Display is a technique based on the expression of synthetic proteins in the phage capsid, and is used for selecting ligands of proteins, cells or tissues. We have produced the recombinant metacaspase of Leishmania (L.) amazonensis and employed Phage Display to find peptide ligands of this enzyme. These peptides led to the identification of potential binding proteins of the MCA, such as kinases and kinesin, which provide information about the regulation and control of MCA´s activity. In the future we will test whether peptide activators of MCA nduce apoptosis of the parasite and can be used as drugs for the treatment of leishmaniasis.
 
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Publishing Date
2013-05-29
 
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