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Purification and characterization of vitellin from the palo verde beetle, Derobracus geminatus

Published online by Cambridge University Press:  19 September 2011

Ellie O. Osir  and
John H. Law
Ellie O. Osir
Affiliation:
Department of Biochemistry, Biological Sciences West, University of Arizona Tucson, AZ 85721, USA
John H. Law
Affiliation:
Department of Biochemistry, Biological Sciences West, University of Arizona Tucson, AZ 85721, USA
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Abstract

Isolation of vitellin from mature eggs of the palo verde beetle, Derobracus geminatus, was achieved by a combination of gel permeation and anion-exchange chromatography. It is a phosphoglycolipoprotein (Mr ˜250,000) that contains 8% lipids, 4% carbohydrates and 0.2% protein-bound phosphorus. The vitellin is composed of three apoproteins designated I (Mr ˜160,000), II (Mr ˜140,000) and III (Mr ˜50,000). The carbohydrate moiety is composed exclusively of mannose and N-acetylglucosamine. All three apoproteins are glycosylated as shown by flourescein isothiocyanate-conjugated concanavalin A and periodate-Schiff (PAS) staining suggesting the presence of a high mannose structure. Further evidence for the high mannose structure was obtained by the ability of endo-β-N-acetylglucosaminidase H treatment to remove the carbohydrate moieties on the apoproteins.

Résumé

L'isolation du vitellus des oeufs murs du Derobracus geminatus fut realisée grâce à une combination de la filtration à travers un colloïde et à la Chromatographie à echange d' ions. C'est une phospholipoproteine (poids moleculaire 250 000) qui contient 8% de lipides, 4% de glucides et 0.2 % de proteines denommees I (poids moleculaie 160 000), II (poids moleculaire 140 000) et III (poids moleculaire 50 000). La fraction de glucides est composée exclusivement de mannose et N-acetylglucosamine. Toutes les 3 apoproteines étaient glycosidees et demontraient la presence de la mannose comme ça etait demontré grâce à la fluorescence du isothiocyanate conjugué à la concanavalin. A et à la coloration au Periodate de Schiff. Une autre evidence pour demontrer l'existence de la mannose fut obtenue grâce à la l'abilité du endo-β-N-acetylglucosaminidase H de detacher les fractions de glucides des apoproteines.

Keywords

Derobracus geminatusvitellinphysical-chemical studiesDerobracus geminatusvitellusetudes physicochimiques
Type
Research Articles
Information
International Journal of Tropical Insect Science , Volume 10 , Issue 3 , June 1989 , pp. 259 - 267
Copyright
Copyright © ICIPE 1989

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