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CHARACTERIZATION OF GLYCOSIDE HYDROLASE FAMILIES 13 AND 31 REVEALS EXPANSION AND DIVERSIFICATION OF α-AMYLASE GENES IN THE PHLEBOTOMINE LUTZOMYIA LONGIPALPIS AND MODULATION OF SANDFLY GLYCOSIDASE ACTIVITIES BY LEISHMANIA INFECTION
Açucar
Sucrase
α-glucosidase
α-amylase
Hidrolase gllicoside
Leishmania mexicana
Sugar
Sucrase
α-glucosidase
α-amylase
glycoside hydrolase
Leishmania mexicana
Affilliation
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil.
Faculty of Health and Medicine, Division of Biomedical and Life Sciences, Lancaster University. Lancaster, United Kingdom.
Faculty of Health and Medicine, Division of Biomedical and Life Sciences, Lancaster University. Lancaster, United Kingdom / Instituto Nacional de Ciência e Tecnologia para Entomologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia para Entomologia Molecular. Rio de Janeiro, RJ, Brasil.
Faculty of Health and Medicine, Division of Biomedical and Life Sciences, Lancaster University. Lancaster, United Kingdom.
Faculty of Health and Medicine, Division of Biomedical and Life Sciences, Lancaster University. Lancaster, United Kingdom / Instituto Nacional de Ciência e Tecnologia para Entomologia Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia para Entomologia Molecular. Rio de Janeiro, RJ, Brasil.
Abstract
Sugar-rich food sources are essential for sandflies to meet their energy demands,
achieving more prolonged survival. The digestion of carbohydrates from food is mainly
realized by glycoside hydrolases (GH). To identify genes coding for α-glycosidases
and α-amylases belonging to Glycoside Hydrolase Family 13 (GH13) and Glycoside
Hydrolase Family 31 (GH31) in Lutzomyia longipalpis, we performed an HMMER search
against its genome using known sequences from other dipteran species. The sequences
retrieved were classified based on BLASTP best hit, analysis of conserved regions
by alignment with sequences of proteins with known structure, and phylogenetic
analysis comparing with orthologous proteins from other dipteran species. Using RT PCR analysis, we evaluated the expression of GH13 and GH31 genes, in the gut and
rest of the body of females, in four different conditions: non-fed, sugar-fed, blood-fed,
and Leishmania mexicana infected females. L. longipalpis has GH13/31 genes that code
for enzymes involved in various aspects of sugar metabolism, as carbohydrate digestion,
storage, and mobilization of glycogen reserves, proteins involved in transport, control
of N-glycosylation quality, as well as others with a putative function in the regulation
of myogenesis. These proteins are representatives of GH13 and GH31 families, and
their roles seem to be conserved. Most of the enzymes seem to be active with
conserved consense sequences, including the expected catalytic residues. α-amylases
also demonstrated the presence of calcium and chloride binding sites. L. longipalpis
genome shows an expansion in the α-amylase gene family, with two clusters. In
contrast, a retraction in the number of α-glucosidases occurred. The expansion of α-amylases is probably related to the specialization of these proteins for different
substrates or inhibitors, which might correlate with the higher diversity of plant foods
available in the natural habitat of L. longipalpis. The expression of α-glucosidase genes
is higher in blood-fed females, suggesting their role in blood digestion. Besides that, in
blood-fed females infected with the parasite Leishmania mexicana, these genes were
also modulated. Glycoside Hydrolases from families 13 and 31 are essential for the
metabolism of L. longipalpis, and GH13 enzymes seem to be involved in the interaction
between sandflies and Leishmania.
Keywords in Portuguese
Lutzomyia longipalpisAçucar
Sucrase
α-glucosidase
α-amylase
Hidrolase gllicoside
Leishmania mexicana
Keywords
Lutzomyia longipalpisSugar
Sucrase
α-glucosidase
α-amylase
glycoside hydrolase
Leishmania mexicana
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