Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display
Autor(a) principal: | |
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Data de Publicação: | 2007 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/15810 |
Resumo: | The genus Dinoponera is represented by six species, geographically distributed along South América. The species Dinoponera australis is found from Mato Grosso to Rio Grande do Sul, as well as in neighboring countries. These ants are characterized by the presence of venom glands that are responsible for the production of a toxin constituted in its majority by proteins. This investigation aimed: (a) the identification of specific ligand peptides to the D. australis venom through phage display, (b) to test the hemolytic activity inhibition by selected phages conjugated to the toxin, (c) to analyse the nucleolytic activity of the venom, and (d) to demonstrate if phage particles are inactivated by the venom. Two commercial random peptide libraries were used: Ph.D.-7 Heptapeptide Phage Display Library and Ph.D.-12 Peptide 12-mer Phage Display Library. After three rounds of selection, 96 phage clones were sequenced and translated, generating 40 valid sequences, and 21 distinct peptides. Bioinformatic analyses have identified three small frequent motifs: KVWxL, WxLL and KLxTIPM. Similarity tests have found as putative human cellular targets acting as: receptors, transcritpion factors, proteins associated with inflamation processes, calcium channel and G protein receptor. In insects, the closest identities found with putative targets were: potassium channel, cytochrome P450, ATPase, transcription factors, and proteinase. The hemolytic activity assays with selected phages conjugated with the venom were all positives, meaning that none of the peptides were able to influence the toxin activity. The nucleolytic activity assay with venom concentrations equal to 2.5 μg was shown by a smeared DNA profile in agarose gel electrophoresis, which may indicate a possible DNA degradation. The crude venom concentration of 10 μg was not able to inactivate phage particles. We have successfully identified interesting peptides that bind specifically to the D. australis venom through Phage Display, which may be important target molecules for the development of therapeutic strategies. |
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2016-06-22T18:43:41Z2009-06-012016-06-22T18:43:41Z2007-05-08SIQUIEROLI, Ana Carolina Silva. Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display. 2007. 77 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2007.https://repositorio.ufu.br/handle/123456789/15810The genus Dinoponera is represented by six species, geographically distributed along South América. The species Dinoponera australis is found from Mato Grosso to Rio Grande do Sul, as well as in neighboring countries. These ants are characterized by the presence of venom glands that are responsible for the production of a toxin constituted in its majority by proteins. This investigation aimed: (a) the identification of specific ligand peptides to the D. australis venom through phage display, (b) to test the hemolytic activity inhibition by selected phages conjugated to the toxin, (c) to analyse the nucleolytic activity of the venom, and (d) to demonstrate if phage particles are inactivated by the venom. Two commercial random peptide libraries were used: Ph.D.-7 Heptapeptide Phage Display Library and Ph.D.-12 Peptide 12-mer Phage Display Library. After three rounds of selection, 96 phage clones were sequenced and translated, generating 40 valid sequences, and 21 distinct peptides. Bioinformatic analyses have identified three small frequent motifs: KVWxL, WxLL and KLxTIPM. Similarity tests have found as putative human cellular targets acting as: receptors, transcritpion factors, proteins associated with inflamation processes, calcium channel and G protein receptor. In insects, the closest identities found with putative targets were: potassium channel, cytochrome P450, ATPase, transcription factors, and proteinase. The hemolytic activity assays with selected phages conjugated with the venom were all positives, meaning that none of the peptides were able to influence the toxin activity. The nucleolytic activity assay with venom concentrations equal to 2.5 μg was shown by a smeared DNA profile in agarose gel electrophoresis, which may indicate a possible DNA degradation. The crude venom concentration of 10 μg was not able to inactivate phage particles. We have successfully identified interesting peptides that bind specifically to the D. australis venom through Phage Display, which may be important target molecules for the development of therapeutic strategies.O gênero Dinoponera apresenta seis espécies, todas com distribuição pela América do Sul, sendo que a Dinoponera australis é encontrada do Mato Grosso até o Rio Grande do Sul e países vizinhos. Essas formigas são caracterizadas pela presença de uma glândula de veneno que é responsável pela produção de uma toxina, constituída em sua maior parte por proteínas. Este trabalho teve como objetivo: (a) identificação de peptídeos ligantes específicos ao veneno de D. australis por Phage Display, (b) testar a inibição da atividade hemolítica pelos fagos selecionados conjugados com o veneno, (c) analisar a atividade nucleásica do veneno e (d) demonstrar se as partículas de fago são inativadas pelo veneno. Duas bibliotecas comerciais de peptídeos randômicos foram utilizadas: Ph.D.-7 Heptapeptide Phage Display Library e Ph.D.-12 Peptide 12-mer Phage Display Library. Após três etapas de seleção, 96 clones tiveram sua seqüência de DNA determinada e foram traduzidos, gerando 40 sequências válidas e 21 peptídeos distintos. As análises de bioinformática identificaram 3 pequenos motivos proteicos: KVWxL, WxLL e KLxTIPM. A análise de similaridade encontrou como prováveis alvos em Humanos receptores celulares, fatores de transcrição, proteínas relacionadas a processos inflamatórios, canal de cálcio e receptor de proteína G. Já em insetos, a análise apontou como possíveis alvos canal de potássio, citocromo, ATPase, fator de transcrição e proteinase. O teste da atividade hemolítica dos fagos selecionados conjugados com veneno foram todos positivos demonstrando que nenhum dos nove peptídeos testados foi capaz de influenciar na ação do veneno. O teste de atividade nucleásica mostrou que nas concentrações de 2,5μg de veneno obteve-se um rastro que indica uma possível degradação do DNA. A concentração de 10μg de veneno bruto não foi capaz de inativar as partículas de fago. Tivemos successo na identificação de peptídeos de interesse, ligantes específicos ao veneno de D. australis por Phage Display, os quais podem ser moléculas alvo importantes para o desenvolvimento de estratégias terapêuticas.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorMestre em Genética e Bioquímicaapplication/pdfporUniversidade Federal de UberlândiaPrograma de Pós-graduação em Genética e BioquímicaUFUBRCiências BiológicasVenenoDinoponera australisFormigaPeptídiosPhage displayVenomCNPQ::CIENCIAS BIOLOGICAS::GENETICAIdentificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage displayinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisGoulart Filho, Luiz Ricardohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781012P8Bonetti, Ana Mariahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783660P9Bacci Junior, Mauriciohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785828J1Bueno, Odair Correahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783063J0http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4779226H8Siquieroli, Ana Carolina Silvainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFUTHUMBNAILAna Maria.pdf.jpgAna Maria.pdf.jpgGenerated Thumbnailimage/jpeg1392https://repositorio.ufu.br/bitstream/123456789/15810/3/Ana%20Maria.pdf.jpgce1169ef0e071e679e8ccdf05bc9cf1fMD53ORIGINALAna Maria.pdfapplication/pdf573457https://repositorio.ufu.br/bitstream/123456789/15810/1/Ana%20Maria.pdfd0f210823779bec733778da93b4402a9MD51TEXTAna Maria.pdf.txtAna Maria.pdf.txtExtracted texttext/plain103826https://repositorio.ufu.br/bitstream/123456789/15810/2/Ana%20Maria.pdf.txt9c00e2ec3cae33482703ec9fd9c2602fMD52123456789/158102016-06-23 04:20:37.438oai:repositorio.ufu.br:123456789/15810Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2016-06-23T07:20:37Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.por.fl_str_mv |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
title |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
spellingShingle |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display Siquieroli, Ana Carolina Silva Veneno Dinoponera australis Formiga Peptídios Phage display Venom CNPQ::CIENCIAS BIOLOGICAS::GENETICA |
title_short |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
title_full |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
title_fullStr |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
title_full_unstemmed |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
title_sort |
Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display |
author |
Siquieroli, Ana Carolina Silva |
author_facet |
Siquieroli, Ana Carolina Silva |
author_role |
author |
dc.contributor.advisor-co1.fl_str_mv |
Goulart Filho, Luiz Ricardo |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781012P8 |
dc.contributor.advisor1.fl_str_mv |
Bonetti, Ana Maria |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783660P9 |
dc.contributor.referee1.fl_str_mv |
Bacci Junior, Mauricio |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785828J1 |
dc.contributor.referee2.fl_str_mv |
Bueno, Odair Correa |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783063J0 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4779226H8 |
dc.contributor.author.fl_str_mv |
Siquieroli, Ana Carolina Silva |
contributor_str_mv |
Goulart Filho, Luiz Ricardo Bonetti, Ana Maria Bacci Junior, Mauricio Bueno, Odair Correa |
dc.subject.por.fl_str_mv |
Veneno Dinoponera australis Formiga Peptídios |
topic |
Veneno Dinoponera australis Formiga Peptídios Phage display Venom CNPQ::CIENCIAS BIOLOGICAS::GENETICA |
dc.subject.eng.fl_str_mv |
Phage display Venom |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::GENETICA |
description |
The genus Dinoponera is represented by six species, geographically distributed along South América. The species Dinoponera australis is found from Mato Grosso to Rio Grande do Sul, as well as in neighboring countries. These ants are characterized by the presence of venom glands that are responsible for the production of a toxin constituted in its majority by proteins. This investigation aimed: (a) the identification of specific ligand peptides to the D. australis venom through phage display, (b) to test the hemolytic activity inhibition by selected phages conjugated to the toxin, (c) to analyse the nucleolytic activity of the venom, and (d) to demonstrate if phage particles are inactivated by the venom. Two commercial random peptide libraries were used: Ph.D.-7 Heptapeptide Phage Display Library and Ph.D.-12 Peptide 12-mer Phage Display Library. After three rounds of selection, 96 phage clones were sequenced and translated, generating 40 valid sequences, and 21 distinct peptides. Bioinformatic analyses have identified three small frequent motifs: KVWxL, WxLL and KLxTIPM. Similarity tests have found as putative human cellular targets acting as: receptors, transcritpion factors, proteins associated with inflamation processes, calcium channel and G protein receptor. In insects, the closest identities found with putative targets were: potassium channel, cytochrome P450, ATPase, transcription factors, and proteinase. The hemolytic activity assays with selected phages conjugated with the venom were all positives, meaning that none of the peptides were able to influence the toxin activity. The nucleolytic activity assay with venom concentrations equal to 2.5 μg was shown by a smeared DNA profile in agarose gel electrophoresis, which may indicate a possible DNA degradation. The crude venom concentration of 10 μg was not able to inactivate phage particles. We have successfully identified interesting peptides that bind specifically to the D. australis venom through Phage Display, which may be important target molecules for the development of therapeutic strategies. |
publishDate |
2007 |
dc.date.issued.fl_str_mv |
2007-05-08 |
dc.date.available.fl_str_mv |
2009-06-01 2016-06-22T18:43:41Z |
dc.date.accessioned.fl_str_mv |
2016-06-22T18:43:41Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
SIQUIEROLI, Ana Carolina Silva. Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display. 2007. 77 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2007. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufu.br/handle/123456789/15810 |
identifier_str_mv |
SIQUIEROLI, Ana Carolina Silva. Identificação de peptídeos ligantes ao veneno de Dinoponera australis (Hymenoptera, Formicidae, Ponerinae) por phage display. 2007. 77 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2007. |
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https://repositorio.ufu.br/handle/123456789/15810 |
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info:eu-repo/semantics/openAccess |
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Universidade Federal de Uberlândia |
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Programa de Pós-graduação em Genética e Bioquímica |
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UFU |
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BR |
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Ciências Biológicas |
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Universidade Federal de Uberlândia |
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