Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Outros Autores: | , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0006966 http://hdl.handle.net/11449/71145 |
Resumo: | Background: Hematophagous insects digest large amounts of host hemoglobin and release heme inside their guts. In Rhodnius prolixus, hemoglobin-derived heme is detoxified by biomineralization, forming hemozoin (Hz). Recently, the involvement of the R. prolixus perimicrovillar membranes in Hz formation was demonstrated. Methodology/Principal Findings: Hz formation activity of an α-glucosidase was investigated. Hz formation was inhibited by specific α-glucosidase inhibitors. Moreover, Hz formation was sensitive to inhibition by Diethypyrocarbonate, suggesting a critical role of histidine residues in enzyme activity. Additionally, a polyclonal antibody raised against a phytophagous insect α-glucosidase was able to inhibit Hz formation. The α-glucosidase inhibitors have had no effects when used 10 h after the start of reaction, suggesting that α-glucosidase should act in the nucleation step of Hz formation. Hz formation was seen to be dependent on the substrate-binding site of enzyme, in a way that maltose, an enzyme substrate, blocks such activity. dsRNA, constructed using the sequence of α-glucosidase gene, was injected into R. prolixus females' hemocoel. Gene silencing was accomplished by reduction of both α-glucosidase and Hz formation activities. Insects were fed on plasma or hemin-enriched plasma and gene expression and activity of α-glucosidase were higher in the plasma plus hemin-fed insects. The deduced amino acid sequence of α-glucosidase shows a high similarity to the insect α-glucosidases, with critical histidine and aspartic residues conserved among the enzymes. Conclusions/Significance: Herein the Hz formation is shown to be associated to an a-glucosidase, the biochemical marker from Hemipteran perimicrovillar membranes. Usually, these enzymes catalyze the hydrolysis of glycosidic bond. The results strongly suggest that α-glucosidase is responsible for Hz nucleation in the R. prolixus midgut, indicating that the plasticity of this enzyme may play an important role in conferring fitness to hemipteran hematophagy, for instance. © 2009 Mury et al. |
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Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary historyalpha glucosidaseaspartic aciddiethyl pyrocarbonatedouble stranded DNAheminhemozoinhistidinemaltosepolyclonal antibodydouble stranded RNAhemehemoglobinhemoproteinamino acid sequenceanimal tissuebinding sitecontrolled studyenzyme activityenzyme bindingenzyme substrategene expressiongene sequencegene silencinginsectnonhumannucleotide sequenceRhodniusrhodnius prolixussuckinganimalcatalysischemistryfemalegene expression regulationhydrolysisintestinemetabolismmicrovillusmolecular evolutionHexapodaRhodnius prolixusalpha-GlucosidasesAnimalsBinding SitesCatalysisEvolution, MolecularFemaleGene Expression RegulationHemeHemeproteinsHemoglobinsHydrolysisInsectsIntestinesMicrovilliRNA, Double-StrandedBackground: Hematophagous insects digest large amounts of host hemoglobin and release heme inside their guts. In Rhodnius prolixus, hemoglobin-derived heme is detoxified by biomineralization, forming hemozoin (Hz). Recently, the involvement of the R. prolixus perimicrovillar membranes in Hz formation was demonstrated. Methodology/Principal Findings: Hz formation activity of an α-glucosidase was investigated. Hz formation was inhibited by specific α-glucosidase inhibitors. Moreover, Hz formation was sensitive to inhibition by Diethypyrocarbonate, suggesting a critical role of histidine residues in enzyme activity. Additionally, a polyclonal antibody raised against a phytophagous insect α-glucosidase was able to inhibit Hz formation. The α-glucosidase inhibitors have had no effects when used 10 h after the start of reaction, suggesting that α-glucosidase should act in the nucleation step of Hz formation. Hz formation was seen to be dependent on the substrate-binding site of enzyme, in a way that maltose, an enzyme substrate, blocks such activity. dsRNA, constructed using the sequence of α-glucosidase gene, was injected into R. prolixus females' hemocoel. Gene silencing was accomplished by reduction of both α-glucosidase and Hz formation activities. Insects were fed on plasma or hemin-enriched plasma and gene expression and activity of α-glucosidase were higher in the plasma plus hemin-fed insects. The deduced amino acid sequence of α-glucosidase shows a high similarity to the insect α-glucosidases, with critical histidine and aspartic residues conserved among the enzymes. Conclusions/Significance: Herein the Hz formation is shown to be associated to an a-glucosidase, the biochemical marker from Hemipteran perimicrovillar membranes. Usually, these enzymes catalyze the hydrolysis of glycosidic bond. The results strongly suggest that α-glucosidase is responsible for Hz nucleation in the R. prolixus midgut, indicating that the plasticity of this enzyme may play an important role in conferring fitness to hemipteran hematophagy, for instance. © 2009 Mury et al.Laboratório de Química e Função de Proteínas e Peptídeos Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de JaneiroLaboratório de Biotecnologia Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de JaneiroInstituto de Química Departamento de Bioquímica and NUPEM Universidade Federal do Rio de Janeiro, Macaé, Rio de JaneiroInstituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de JaneiroDepartamento de Parasitologia Universidade Estadual de São Paulo, Botucatu, São PauloDepartamento de Bioquímica Universidade Federal de Santa Catarina, Florianópolis, Santa CatarinaDepartment of Molecular Microbiology and Immunology Johns Hopkins University Bloomberg School of Public Health, Baltimore, MDDepartamento de Parasitologia Universidade Estadual de São Paulo, Botucatu, São PauloUniversidade Estadual do Norte FluminenseUniversidade Federal do Rio de Janeiro (UFRJ)Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM)Universidade Estadual Paulista (Unesp)Universidade Federal de Santa Catarina (UFSC)Bloomberg School of Public HealthMury, Flávia Borgesda Silva, José RobertoFerreira, Ligia Souzados Santos Ferreira, Beatrizde Souza-Filho, Gonçalo Apolináriode Souza-Neto, Jayme AugustoRibolla, Paulo Eduardo Martins [UNESP]Silva, Carlo Peresdo Nascimento, Viviane VeigaMachado, Olga Lima TavaresBerbert-Molina, Morilla AmorimDansa-Petretski, Marilvia2014-05-27T11:23:58Z2014-05-27T11:23:58Z2009-09-09info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1371/journal.pone.0006966PLoS ONE, v. 4, n. 9, 2009.1932-6203http://hdl.handle.net/11449/7114510.1371/journal.pone.00069662-s2.0-703491308012-s2.0-70349130801.pdf35771497484568800000-0001-8735-6090Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPLOS ONE2.7661,164info:eu-repo/semantics/openAccess2023-10-26T06:06:16Zoai:repositorio.unesp.br:11449/71145Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-26T06:06:16Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
title |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
spellingShingle |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history Mury, Flávia Borges alpha glucosidase aspartic acid diethyl pyrocarbonate double stranded DNA hemin hemozoin histidine maltose polyclonal antibody double stranded RNA heme hemoglobin hemoprotein amino acid sequence animal tissue binding site controlled study enzyme activity enzyme binding enzyme substrate gene expression gene sequence gene silencing insect nonhuman nucleotide sequence Rhodnius rhodnius prolixus sucking animal catalysis chemistry female gene expression regulation hydrolysis intestine metabolism microvillus molecular evolution Hexapoda Rhodnius prolixus alpha-Glucosidases Animals Binding Sites Catalysis Evolution, Molecular Female Gene Expression Regulation Heme Hemeproteins Hemoglobins Hydrolysis Insects Intestines Microvilli RNA, Double-Stranded |
title_short |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
title_full |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
title_fullStr |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
title_full_unstemmed |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
title_sort |
Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history |
author |
Mury, Flávia Borges |
author_facet |
Mury, Flávia Borges da Silva, José Roberto Ferreira, Ligia Souza dos Santos Ferreira, Beatriz de Souza-Filho, Gonçalo Apolinário de Souza-Neto, Jayme Augusto Ribolla, Paulo Eduardo Martins [UNESP] Silva, Carlo Peres do Nascimento, Viviane Veiga Machado, Olga Lima Tavares Berbert-Molina, Morilla Amorim Dansa-Petretski, Marilvia |
author_role |
author |
author2 |
da Silva, José Roberto Ferreira, Ligia Souza dos Santos Ferreira, Beatriz de Souza-Filho, Gonçalo Apolinário de Souza-Neto, Jayme Augusto Ribolla, Paulo Eduardo Martins [UNESP] Silva, Carlo Peres do Nascimento, Viviane Veiga Machado, Olga Lima Tavares Berbert-Molina, Morilla Amorim Dansa-Petretski, Marilvia |
author2_role |
author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual do Norte Fluminense Universidade Federal do Rio de Janeiro (UFRJ) Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM) Universidade Estadual Paulista (Unesp) Universidade Federal de Santa Catarina (UFSC) Bloomberg School of Public Health |
dc.contributor.author.fl_str_mv |
Mury, Flávia Borges da Silva, José Roberto Ferreira, Ligia Souza dos Santos Ferreira, Beatriz de Souza-Filho, Gonçalo Apolinário de Souza-Neto, Jayme Augusto Ribolla, Paulo Eduardo Martins [UNESP] Silva, Carlo Peres do Nascimento, Viviane Veiga Machado, Olga Lima Tavares Berbert-Molina, Morilla Amorim Dansa-Petretski, Marilvia |
dc.subject.por.fl_str_mv |
alpha glucosidase aspartic acid diethyl pyrocarbonate double stranded DNA hemin hemozoin histidine maltose polyclonal antibody double stranded RNA heme hemoglobin hemoprotein amino acid sequence animal tissue binding site controlled study enzyme activity enzyme binding enzyme substrate gene expression gene sequence gene silencing insect nonhuman nucleotide sequence Rhodnius rhodnius prolixus sucking animal catalysis chemistry female gene expression regulation hydrolysis intestine metabolism microvillus molecular evolution Hexapoda Rhodnius prolixus alpha-Glucosidases Animals Binding Sites Catalysis Evolution, Molecular Female Gene Expression Regulation Heme Hemeproteins Hemoglobins Hydrolysis Insects Intestines Microvilli RNA, Double-Stranded |
topic |
alpha glucosidase aspartic acid diethyl pyrocarbonate double stranded DNA hemin hemozoin histidine maltose polyclonal antibody double stranded RNA heme hemoglobin hemoprotein amino acid sequence animal tissue binding site controlled study enzyme activity enzyme binding enzyme substrate gene expression gene sequence gene silencing insect nonhuman nucleotide sequence Rhodnius rhodnius prolixus sucking animal catalysis chemistry female gene expression regulation hydrolysis intestine metabolism microvillus molecular evolution Hexapoda Rhodnius prolixus alpha-Glucosidases Animals Binding Sites Catalysis Evolution, Molecular Female Gene Expression Regulation Heme Hemeproteins Hemoglobins Hydrolysis Insects Intestines Microvilli RNA, Double-Stranded |
description |
Background: Hematophagous insects digest large amounts of host hemoglobin and release heme inside their guts. In Rhodnius prolixus, hemoglobin-derived heme is detoxified by biomineralization, forming hemozoin (Hz). Recently, the involvement of the R. prolixus perimicrovillar membranes in Hz formation was demonstrated. Methodology/Principal Findings: Hz formation activity of an α-glucosidase was investigated. Hz formation was inhibited by specific α-glucosidase inhibitors. Moreover, Hz formation was sensitive to inhibition by Diethypyrocarbonate, suggesting a critical role of histidine residues in enzyme activity. Additionally, a polyclonal antibody raised against a phytophagous insect α-glucosidase was able to inhibit Hz formation. The α-glucosidase inhibitors have had no effects when used 10 h after the start of reaction, suggesting that α-glucosidase should act in the nucleation step of Hz formation. Hz formation was seen to be dependent on the substrate-binding site of enzyme, in a way that maltose, an enzyme substrate, blocks such activity. dsRNA, constructed using the sequence of α-glucosidase gene, was injected into R. prolixus females' hemocoel. Gene silencing was accomplished by reduction of both α-glucosidase and Hz formation activities. Insects were fed on plasma or hemin-enriched plasma and gene expression and activity of α-glucosidase were higher in the plasma plus hemin-fed insects. The deduced amino acid sequence of α-glucosidase shows a high similarity to the insect α-glucosidases, with critical histidine and aspartic residues conserved among the enzymes. Conclusions/Significance: Herein the Hz formation is shown to be associated to an a-glucosidase, the biochemical marker from Hemipteran perimicrovillar membranes. Usually, these enzymes catalyze the hydrolysis of glycosidic bond. The results strongly suggest that α-glucosidase is responsible for Hz nucleation in the R. prolixus midgut, indicating that the plasticity of this enzyme may play an important role in conferring fitness to hemipteran hematophagy, for instance. © 2009 Mury et al. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-09-09 2014-05-27T11:23:58Z 2014-05-27T11:23:58Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1371/journal.pone.0006966 PLoS ONE, v. 4, n. 9, 2009. 1932-6203 http://hdl.handle.net/11449/71145 10.1371/journal.pone.0006966 2-s2.0-70349130801 2-s2.0-70349130801.pdf 3577149748456880 0000-0001-8735-6090 |
url |
http://dx.doi.org/10.1371/journal.pone.0006966 http://hdl.handle.net/11449/71145 |
identifier_str_mv |
PLoS ONE, v. 4, n. 9, 2009. 1932-6203 10.1371/journal.pone.0006966 2-s2.0-70349130801 2-s2.0-70349130801.pdf 3577149748456880 0000-0001-8735-6090 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
PLOS ONE 2.766 1,164 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1799964702969692160 |