Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells

Andrii Kaberniuk, A. J. Labyntsev, D. V. Kolibo, O. S. Oliinyk, T. A. Redchuk, N. V. Korotkevich, V. F. Gorchev, S. O. Karahim, S. V. Komisarenko

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)67-77
Number of pages11
JournalUkrain'skyi Biokhimichnyi Zhurnal
Volume81
Issue number1
StatePublished - Apr 27 2009
Externally publishedYes

Fingerprint

Diphtheria Toxin
Vero Cells
Derivatives
Flow cytometry
Fluorescein-5-isothiocyanate
Confocal microscopy
Fluorescein
Labels
Fusion reactions
Endosomes
Cell Surface Receptors
Confocal Microscopy
Flow Cytometry
Heparin-binding EGF-like Growth Factor
enhanced green fluorescent protein
isothiocyanic acid

Keywords

  • Diphtheria toxin
  • Diphtheria toxin receptor interaction
  • Diphtheria toxin's fluorescent derivatives
  • Fluorescent probes
  • Green fluorescent protein HB-EGF
  • Recombinant subunit B of diphtheria toxin

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kaberniuk, A., Labyntsev, A. J., Kolibo, D. V., Oliinyk, O. S., Redchuk, T. A., Korotkevich, N. V., ... Komisarenko, S. V. (2009). Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells. Ukrain'skyi Biokhimichnyi Zhurnal, 81(1), 67-77.

Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells. / Kaberniuk, Andrii; 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.

Research output: Contribution to journalArticle

Kaberniuk, A, Labyntsev, AJ, Kolibo, DV, Oliinyk, OS, Redchuk, TA, Korotkevich, NV, Gorchev, VF, Karahim, SO & Komisarenko, SV 2009, 'Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells', Ukrain'skyi Biokhimichnyi Zhurnal, vol. 81, no. 1, pp. 67-77.
Kaberniuk A, Labyntsev AJ, Kolibo DV, Oliinyk OS, Redchuk TA, Korotkevich NV et al. Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells. Ukrain'skyi Biokhimichnyi Zhurnal. 2009 Apr 27;81(1):67-77.
Kaberniuk, Andrii ; Labyntsev, A. J. ; Kolibo, D. V. ; Oliinyk, O. S. ; Redchuk, T. A. ; Korotkevich, N. V. ; Gorchev, V. F. ; Karahim, S. O. ; Komisarenko, S. V. / Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells. In: Ukrain'skyi Biokhimichnyi Zhurnal. 2009 ; Vol. 81, No. 1. pp. 67-77.
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