Diphtheria toxin's B subunit provides toxin interaction with its receptor on the cell surface and translocation of toxin's A subunit from endosome to cytozole of sensitive cells. Functional analogues of B subunit with fluorescent label are considered as perspective tools for studying the above mentioned processes. The aim of the work was to obtain fluorescent B subunit analogues and to detect the specificity of their interaction with Vero line cells. B subunit fluorescent analogues were obtained in two different ways. The first one was B subunit chemical conjugation with fluorescein isothiocyanate and the second one was genetic fusion of recombinant B subunit chain with enhanced green fluorescent protein chain. Specific interaction of B subunit fluorescent derivatives with Vero cells was studied by flow cytometry and confocal microscopy. Using competitive analysis it was shown that B subunit fluorescent analogues possessed different affinity for cells. The affinity of EGFP-SbB was higher than FITC-SbB. Our results indicate the possibility to use the fluorescent derivatives of B subunit as tools for identification of diphtheria toxin's receptor (HB-EGF) expression on the cell surface as well as for studying the interaction and penetration of diphtheria toxin to the cell.
Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells. / Kabemiuk, A. A.; Labyntsev, A. J.; Kolibo, D. V.; Oliinyk, O. S.; Redchuk, T. A.; Korotkevich, N. V.; Gorchev, V. F.; Karahim, S. O.; Komisarenko, S. V.In: Ukrain'skyi Biokhimichnyi Zhurnal, Vol. 81, No. 1, 27.04.2009, p. 67-77.
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