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Isolation and properties of a 23-KD haemolymph protein from the tsetse, Glossina morsitans morsitans
Published online by Cambridge University Press: 19 September 2011
Abstract
The haemolymph of the tsetse, Glossina morsitans morsitans, contains a low molecular weight protein of very high density (1.29 g/mi). The protein was detected in the haemolymph during all developmental stages of the insect. Purification of the protein was achieved by a combination of density gradient ultracentrifugation and repeated gel permeation chromatography. Electrophoresis under non-denaturing and denaturing conditions showed the protein to be a single polypeptide chain (M,˜23,000). Amino acid analysis revealed a relatively high content of the acidic amino acids as well as serine and glycine. The protein contained lipids as shown by Sudan Black staining but was nonglycosylated. Using rabbit antiserum against the isolated protein in immunodiffusion and immunoblotting experiments, no cross-reactivity was detected with haemolymph samples from insects representing six orders. Although the function of the protein remains unknown, its uniqueness to Glossina suggests that it may have a role in the physiology of this insect.
Résumé
L'hémolymphe de la mouche tsetsé, Glossina morsitans morsitans, renferme une protéine de faible poids moléculaire à densité très élevée (1.29 g/ml). La protéin a été décelée dans l'hémolymphe à tous les stades de développement de l'insecte. Sa purification est obtenu grâce à une combinaison de I'ultracentrifugation par gradient de densité et de la chromatographie repetée sur gel filtrant. L'électrophorèse en conditions non denaturante et denaturante a révelé que la protéine est une simple chaîne de polypeptide (Mr = 23,000) serine et glycine.
L'analyse des acides aminés a perm is de déceler un taux élevée d'acides aminés acides. La protéine contient également des lipides relevés par coloration au Noir du soudan, mais était non glycolisée. En utilisant de l'antisérum de lapin sur la protéine isolée au cours de l'immunodiffusion et l'immunotampon, aucune réaction croisée fut détectée avec des échantillons d'hémolymphe d'insectes représentant six ordres. La fonction de la protéin reste cependant inconnue mais sa spécificité par rapport au Glossina, laisse supposer qu'elle pourrait Jouer un rôle important dans la physiologie de cet insecte.
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References
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