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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2009.tde-11022010-104758
Document
Author
Full name
Linda Christian Carrijo Carvalho
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Tavassi, Ana Marisa Chudzinski (President)
Araújo, Heloisa Sobreiro Selistre de
Gonçalves, Viviane Maimoni
Martins, Elizabeth Angelica Leme
Porcionatto, Marimelia Aparecida
Title in Portuguese
LOPAP (Lonomia obliqua prothrombin activator protease): clonagem e expressão em levedura Pichia pastoris, obtenção de um peptídeo sintético, análise estrutural e avaliação de suas potenciais aplicações.
Keywords in Portuguese
Biologia celular
Bioquímica
Hemostasia
Matriz extracelular
Peptídeos
Proteínas recombinantes
Abstract in Portuguese
O Lopap é um ativador de protrombina da lagarta L. obliqua, pertence à família das lipocalinas e apresenta atividade antiapoptótica. O Lopap foi obtido na forma recombinante (rLopap), na levedura P. pastoris, por metodologia escalonável, e sua atividade foi avaliada in vitro e in vivo. O tratamento com rLopap reduziu o tempo de sangramento em animais anticoagulados com enoxaparina. Por outro lado, um peptídeo derivado do Lopap, designado antiapoptotic peptide (AP), foi capaz de induzir a síntese de colágeno em cultura de fibroblastos e na derme de animais. A região correspondente a AP apresentou propriedades físicas e estruturais semelhantes a seqüências relacionadas em outras lipocalinas com atividade antiapoptótica. Estes resultados abrem perspectivas para aplicações do Lopap, como uma molécula procoagulante, e de AP, através de sua ação na modulação celular, como um componente cosmético, no reparo e remodelamento tecidual e em disfunções que envolvem morte celular e perda de colágeno.
Title in English
LOPAP (Lonomia obliqua prothrombin activator protease): cloning and expression in Pichia pastoris yeast, design of a synthetic peptide, structural analysis and evaluation of its potential applications.
Keywords in English
Biochemistry
Cell Biology
Extracellular matrix
Hemostasis
Peptides
Recombinant proteins
Abstract in English
Lopap is a prothrombin activator from the L. obliqua caterpillar, belongs to the lipocalin family, and displays antiapoptotic activity. Lopap was obtained in the recombinant form (rLopap) in the P. pastoris yeast, by a scaled up methodology, and its activity was evaluated in vitro and in vivo. Treatment with rLopap reduced the bleeding time in animals anticoagulated with enoxaparin. On the other hand, a Lopap-derived peptide, designated antiapoptotic peptide (AP), was able to induce collagen synthesis in fibroblast culture and in the animal dermis. The region corresponding to AP had similar physical and structural properties when compared with other antiapoptotic lipocalins. These results open perspectives for the use of Lopap, as a procoagulante molecule, and the use of AP, based on its cell modulation effects, as a cosmetic component, aiding tissue repair and in dysfunctions involving cell death and loss of collagen.
 
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Publishing Date
2010-02-18
 
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