Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history
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Data
2009-09-09
Autores
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
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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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Palavras-chave
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
Como citar
PLoS ONE, v. 4, n. 9, 2009.