Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)

ABSTRACT The present investigation deals with the isolation and characterization of a lectin from Aedes aegypti (Ae aegypti) female mid gut extract that agglutinates various mammalian red blood cells (RBCs) such as human three groups A, B, and O (RH+), mouse, rat, guinea-pig, sheep and goat erythrocytes. The highest activity of both crude and isolated mid gut lectins were detected against sheep RBCs. Using (NH4)2 SO4 fractionation, ion-exchange and mannose-CNBr-Sepharose 6B affinity chromatography techniques, Ae. aegypti midgut lectin (Aelec) was purified to homogeneity.Isoelectric focusing (IEF) and reducing SDS/PAGE revealed that the isolated mid gut lectin had isoelectric point (PI) of 5.90, and subunits approximate molecular weights of 35.50 and 27.35 KDa. The hemagglutination (HA) of lectins were Ca2+ - independent and heat-resistant. The sugar specificity of the purified Aelec was strongly inhibited by D (+)-mannose and raffinose, followed by D (+) glucose. N-acetyl-D-manosamine and N-acetyl-D-glucosamine were moderate inhibitors. None of the lectins were inhibited by the disaccharides such as galactose, lactose, trehalose (IC50 up to 200 mM or fetuin up to 1%) but the glycosubstances mucin and laminarin were strong inhibitors up to very low concentrations (0.030 - 0.003%).

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Main Authors: Ayaad,Tahany Hassan, Al-Akeel,Rasha Khalifah, Olayan,Ebtisam
Format: Digital revista
Language:English
Published: Instituto de Tecnologia do Paraná - Tecpar 2015
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000600905
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spelling oai:scielo:S1516-891320150006009052015-12-01Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)Ayaad,Tahany HassanAl-Akeel,Rasha KhalifahOlayan,Ebtisam Lectin purification characterization hemagglutination Aedes aegypti mosquitoes ABSTRACT The present investigation deals with the isolation and characterization of a lectin from Aedes aegypti (Ae aegypti) female mid gut extract that agglutinates various mammalian red blood cells (RBCs) such as human three groups A, B, and O (RH+), mouse, rat, guinea-pig, sheep and goat erythrocytes. The highest activity of both crude and isolated mid gut lectins were detected against sheep RBCs. Using (NH4)2 SO4 fractionation, ion-exchange and mannose-CNBr-Sepharose 6B affinity chromatography techniques, Ae. aegypti midgut lectin (Aelec) was purified to homogeneity.Isoelectric focusing (IEF) and reducing SDS/PAGE revealed that the isolated mid gut lectin had isoelectric point (PI) of 5.90, and subunits approximate molecular weights of 35.50 and 27.35 KDa. The hemagglutination (HA) of lectins were Ca2+ - independent and heat-resistant. The sugar specificity of the purified Aelec was strongly inhibited by D (+)-mannose and raffinose, followed by D (+) glucose. N-acetyl-D-manosamine and N-acetyl-D-glucosamine were moderate inhibitors. None of the lectins were inhibited by the disaccharides such as galactose, lactose, trehalose (IC50 up to 200 mM or fetuin up to 1%) but the glycosubstances mucin and laminarin were strong inhibitors up to very low concentrations (0.030 - 0.003%).info:eu-repo/semantics/openAccessInstituto de Tecnologia do Paraná - TecparBrazilian Archives of Biology and Technology v.58 n.6 20152015-12-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000600905en10.1590/S1516-89132015060277
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countrycode BR
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libraryname SciELO
language English
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author Ayaad,Tahany Hassan
Al-Akeel,Rasha Khalifah
Olayan,Ebtisam
spellingShingle Ayaad,Tahany Hassan
Al-Akeel,Rasha Khalifah
Olayan,Ebtisam
Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)
author_facet Ayaad,Tahany Hassan
Al-Akeel,Rasha Khalifah
Olayan,Ebtisam
author_sort Ayaad,Tahany Hassan
title Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)
title_short Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)
title_full Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)
title_fullStr Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)
title_full_unstemmed Isolation and Characterization of Midgut Lectin From Aedes aegypti (L.) (Diptera: Culicidae)
title_sort isolation and characterization of midgut lectin from aedes aegypti (l.) (diptera: culicidae)
description ABSTRACT The present investigation deals with the isolation and characterization of a lectin from Aedes aegypti (Ae aegypti) female mid gut extract that agglutinates various mammalian red blood cells (RBCs) such as human three groups A, B, and O (RH+), mouse, rat, guinea-pig, sheep and goat erythrocytes. The highest activity of both crude and isolated mid gut lectins were detected against sheep RBCs. Using (NH4)2 SO4 fractionation, ion-exchange and mannose-CNBr-Sepharose 6B affinity chromatography techniques, Ae. aegypti midgut lectin (Aelec) was purified to homogeneity.Isoelectric focusing (IEF) and reducing SDS/PAGE revealed that the isolated mid gut lectin had isoelectric point (PI) of 5.90, and subunits approximate molecular weights of 35.50 and 27.35 KDa. The hemagglutination (HA) of lectins were Ca2+ - independent and heat-resistant. The sugar specificity of the purified Aelec was strongly inhibited by D (+)-mannose and raffinose, followed by D (+) glucose. N-acetyl-D-manosamine and N-acetyl-D-glucosamine were moderate inhibitors. None of the lectins were inhibited by the disaccharides such as galactose, lactose, trehalose (IC50 up to 200 mM or fetuin up to 1%) but the glycosubstances mucin and laminarin were strong inhibitors up to very low concentrations (0.030 - 0.003%).
publisher Instituto de Tecnologia do Paraná - Tecpar
publishDate 2015
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000600905
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