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 language | English (US) |
|---|---|
| Pages (from-to) | 67-77 |
| Number of pages | 11 |
| Journal | Ukrain'skyi Biokhimichnyi Zhurnal |
| Volume | 81 |
| Issue number | 1 |
| State | Published - Apr 27 2009 |
| Externally published | Yes |
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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
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 journal › Article
}
TY - JOUR
T1 - Fluorescent derivatives of diphtheria toxin's subunit B and their interaction with Vero cells
AU - Kaberniuk, Andrii
AU - Labyntsev, A. J.
AU - Kolibo, D. V.
AU - Oliinyk, O. S.
AU - Redchuk, T. A.
AU - Korotkevich, N. V.
AU - Gorchev, V. F.
AU - Karahim, S. O.
AU - Komisarenko, S. V.
PY - 2009/4/27
Y1 - 2009/4/27
N2 - 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.
AB - 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.
KW - Diphtheria toxin
KW - Diphtheria toxin receptor interaction
KW - Diphtheria toxin's fluorescent derivatives
KW - Fluorescent probes
KW - Green fluorescent protein HB-EGF
KW - Recombinant subunit B of diphtheria toxin
UR - http://www.scopus.com/inward/record.url?scp=65249161719&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=65249161719&partnerID=8YFLogxK
M3 - Article
C2 - 19877418
AN - SCOPUS:65249161719
VL - 81
SP - 67
EP - 77
JO - Ukrainian biochemical journal
JF - Ukrainian biochemical journal
SN - 2409-4943
IS - 1
ER -