PHYSICO-CHEMICAL CHARACTERIZATION OF A NATURALLY OCCURRING Ca2+ DEPENDENT MIDGUT GLAND AGGLUTININ IN THE RUSTY MILLIPEDE TRIGONIULUS CORALLINUS
European Journal of Molecular & Clinical Medicine,
2022, Volume 9, Issue 8, Pages 929-942
AbstractObjective: To study the physico-chemical characteristics as well as the binding specificity of the agglutinin in the extract of the midgut gland from the rusty millipede, Trigoniulus corallinus.
Methods: The presence of agglutinin was investigated by hemagglutination assay using mammalian erythrocytes. After selecting the tissue (midgut gland) and the erythrocyte (rabbit) based on high HA titre, the correlation between biochemical parameters of midgut gland agglutinin such as protein, calcium and water content and HA titre was assessed. The influence of physico-chemical parameters such as pH, temperature, divalent cations (Ca2+, Mg2+, Mn2+) and chelators (di- and tetrasodium EDTA and trisodium citrate) on hemagglutinability was tested. The sugar specificity was assessed by hemagglutination inhibition assay with different sugars and glycoproteins.
Results: Physico-chemical analysis of the midgut gland agglutinin exhibited that the activity was optimum at pH 6.5, temperature 35oC and in the presence of calcium ions. The agglutinin activity was sensitive to di and tetra sodium EDTA and trisodium citrate. Hemagglutinability was inhibited by sugars: α-lactose, D-galactosamine, dextrose, GluNAc, D-galactoseand glycoprotein: lactoferrin. A single agglutinin was found when agglutitinability vanished after cross adsorption.
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