Estudos estruturais e de atividades biol?gicas do soyuretox, um pept?deo derivado de urease. Zebrafish (Danio rerio) como modelo de estudo do pept?deo

• Main Author: Kappaun, Karine
• Other Authors: Carlini, C?lia Regina Ribeiro da Silva
• Format: Others
• Language: Portuguese
• Published: Pontif?cia Universidade Cat?lica do Rio Grande do Sul 2018
• Subjects: Soyuretox; Ureases; Estrutura Tridimensional; RMN; Zebrafish; Candida Albicans; Rhodnius Prolixus; Prote?nas Desordenadas; CIENCIAS DA SAUDE::MEDICINA
• Online Access: http://tede2.pucrs.br/tede2/handle/tede/8288

Submitted by PPG Medicina e Ci?ncias da Sa?de ([e-mail]) on 2018-09-17T18:15:32Z No. of bitstreams: 1 KARINE_KAPPAUN.pdf: 4206975 bytes, checksum: b450fa28646271e7dfac9389de975825 (MD5) === Approved for entry into archive by Sheila Dias ([e-mail]) on 2018-09-18T12:20:56Z (GMT) No. of bitstreams: 1 KARINE_KAPPAUN.pdf: 4206975 bytes, checksum: b450fa28646271e7dfac9389de975825 (MD5) === Made available in DSpace on 2018-09-18T12:54:15Z (GMT). No. of bitstreams: 1 KARINE_KAPPAUN.pdf: 4206975 bytes, checksum: b450fa28646271e7dfac9389de975825 (MD5) Previous issue date: 2018-07-06 === Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES === Ureases (urea amidohydrolases, EC 3.5.1.5) are nickel-dependent enzymes, widely spread in bacteria, fungi and plants, which catalyze the hydrolysis of urea to ammonia and carbon dioxide. Jaburetox is a peptide derived from a urease isoform of the Canavalia ensiformis plant. Structurally, jaburetox is an intrinsically disordered peptide, demonstrated by bioinformatics tools, and experimentally, by circular dichroism (CD) and nuclear magnetic resonance (NMR). Several biological properties of jaburetox have already been described, such as insecticidal activity, ability to interact with lipid bilayers, fungitoxic effect, among others. Soyuretox is a peptide colinear to jaburetox derived from the soybean ubiquitous urease. One of the objectives of this thesis was to carry out structural studies of the soyuretox peptide by CD and NMR, which revealed its intrinsically disordered nature and that its secondary structure content is modified in the presence of SDS micelles. Biological properties of soyuretox were evaluated. The peptide has entomotoxic activity, inducing aggregation of hemocytes from Rhodnius prolixus, the vector of Chagas disease, both in vivo and in vitro. The fungitoxic effect of soyuretox on yeast Candida albicans involves the production of superoxide anions, and the peptide was immunolocalized on the surface of the yeast. Finally, the behavioral and morphological effects of soyuretox on zebrafish larvae (Danio rerio) were evaluated, aiming to deepen the understanding of the mechanism of toxic action and the toxicological profile of this molecule, which presents biotechnological potential as a biopesticide. === Ureases (ureia amido-hidrolases; EC 3.5.1.5) s?o enzimas n?quel dependentes, amplamente distribu?das em bact?rias, fungos e plantas, que catalisam a hidr?lise da ureia ? am?nia e di?xido de carbono. O jaburetox ? um pept?deo derivado de uma isoforma de urease da planta Canavalia ensiformis. Estruturalmente, o jaburetox ? um pept?deo intrinsecamente desordenado, demonstrado por ferramentas in silico de bioinform?tica, e experimentalmente, por dicro?smo circular (CD) e resson?ncia magn?tica nuclear (RMN). Diversas propriedades biol?gicas do jaburetox j? foram descritas, tais como atividade inseticida, capacidade de interagir com bicamadas lip?dicas, efeito fungit?xico, dentre outras. O soyuretox ? um pept?deo colinear ao jaburetox, por?m derivado da urease ub?qua de soja. Um dos objetivos dessa tese foi realizar estudos estruturais do pept?deo soyuretox por CD e RMN, que revelaram sua natureza intrinsecamente desordenada e aumento no conte?do de estrutura secund?ria na presen?a de micelas de SDS. Propriedades biol?gicas do soyuretox foram avaliadas. O pept?deo tem atividade entomot?xica, induzindo agrega??o de hem?citos de Rhodnius prolixus, o vetor da doen?a de Chagas, tanto in vivo como in vitro. O efeito fungit?xico do soyuretox na levedura Candida albicans envolve a produ??o de ?nions super?xido, sendo o pept?deo imunolocalizado ligado na superf?cie da levedura. Por fim, foram avaliados os efeitos comportamentais e morfol?gicos do soyuretox em larvas de zebrafish (Danio rerio), visando aprofundar o entendimento do mecanismo de a??o t?xica e o perfil toxicol?gico dessa mol?cula, que apresenta potencial biotecnol?gico como biopesticida.