Effect of the T-domain on intracellular transport of diphtheria toxin
Subunit B of diphtheria toxin (DT), which consists of two domains: R (receptor-binding) and T (transmembrane), plays an important role in toxin-receptor binding on the cell-targets and in transportation of catalytic subunit A to the cell cytosol. Recombinant analogues of the subunit B are promising...
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| Format: | Article |
| Language: | English |
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National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry
2014-06-01
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| Series: | The Ukrainian Biochemical Journal |
| Subjects: | |
| Online Access: | http://ukrbiochemjournal.org/wp-content/uploads/2015/06/Labyntsev_3_14.pdf |
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| author | А. J. Labyntsev D. V. Kolybo E. S. Yurchenko A. A. Kaberniuk N. V. Korotkevych S. V. Komisarenko |
| author_facet | А. J. Labyntsev D. V. Kolybo E. S. Yurchenko A. A. Kaberniuk N. V. Korotkevych S. V. Komisarenko |
| author_sort | А. J. Labyntsev |
| collection | DOAJ |
| description | Subunit B of diphtheria toxin (DT), which consists of two domains: R (receptor-binding) and T (transmembrane), plays an important role in toxin-receptor binding on the cell-targets and in transportation of catalytic subunit A to the cell cytosol. Recombinant analogues of the subunit B are promising representatives in the unique class of transporting proteins, able to deliver different types of biologically active molecules to cell cytosol. In the development of these protein constructs understanding of the role of each of the DT fragments in determination of transporting pathways of endocytosed complex toxin-receptor is urgently required.
We have studied in this work the T-domain effect on intracellular transport of recombinant fragments of DT. We have compared intracellular transport of the R-domain and the subunit B, the last one consisted of both R-domain and T-domain. Recombinant fragments of DT used in this work were labeled with fluorescent proteins, which allowed applying colocalization technique for our study. Application of confocal microscopy technique revealed differences in transportation of recombinant derivates of DT in Vero cells: R domain moved faster than subunit B to tubular compartments. Analysis of R-domain and subunit B transportation confirmed almost linear increase of their colocalization with the time regarding to Pearsons correlation coefficient (PCC). However, amount of colocalized with R-domain subunit B were not linearly increased with time according to Manders coefficient (M1), this could indicate the ability of subunit B to transport to such compartments that R-domain do not reach. Possible role of the T-domain in intracellular transportation and compartmentalization of the toxin may be associated with the ability of the T-domain to form a proton channels and its ability to interact with COPI complex. |
| first_indexed | 2024-03-09T07:30:33Z |
| format | Article |
| id | doaj.art-b834990b7b34416cafa8948298927f43 |
| institution | Directory Open Access Journal |
| issn | 2409-4943 2413-5003 |
| language | English |
| last_indexed | 2024-03-09T07:30:33Z |
| publishDate | 2014-06-01 |
| publisher | National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry |
| record_format | Article |
| series | The Ukrainian Biochemical Journal |
| spelling | doaj.art-b834990b7b34416cafa8948298927f432023-12-03T06:26:36ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryThe Ukrainian Biochemical Journal2409-49432413-50032014-06-01863778710.15407/ubj86.03.077Effect of the T-domain on intracellular transport of diphtheria toxinА. J. Labyntsev0D. V. Kolybo1E. S. Yurchenko2A. A. Kaberniuk3N. V. Korotkevych4S. V. Komisarenko5Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, KyivPalladin Institute of Biochemistry, National Academy of Sciences of Ukraine, KyivPalladin Institute of Biochemistry, National Academy of Sciences of Ukraine, KyivPalladin Institute of Biochemistry, National Academy of Sciences of Ukraine, KyivPalladin Institute of Biochemistry, National Academy of Sciences of Ukraine, KyivPalladin Institute of Biochemistry, National Academy of Sciences of Ukraine, KyivSubunit B of diphtheria toxin (DT), which consists of two domains: R (receptor-binding) and T (transmembrane), plays an important role in toxin-receptor binding on the cell-targets and in transportation of catalytic subunit A to the cell cytosol. Recombinant analogues of the subunit B are promising representatives in the unique class of transporting proteins, able to deliver different types of biologically active molecules to cell cytosol. In the development of these protein constructs understanding of the role of each of the DT fragments in determination of transporting pathways of endocytosed complex toxin-receptor is urgently required. We have studied in this work the T-domain effect on intracellular transport of recombinant fragments of DT. We have compared intracellular transport of the R-domain and the subunit B, the last one consisted of both R-domain and T-domain. Recombinant fragments of DT used in this work were labeled with fluorescent proteins, which allowed applying colocalization technique for our study. Application of confocal microscopy technique revealed differences in transportation of recombinant derivates of DT in Vero cells: R domain moved faster than subunit B to tubular compartments. Analysis of R-domain and subunit B transportation confirmed almost linear increase of their colocalization with the time regarding to Pearsons correlation coefficient (PCC). However, amount of colocalized with R-domain subunit B were not linearly increased with time according to Manders coefficient (M1), this could indicate the ability of subunit B to transport to such compartments that R-domain do not reach. Possible role of the T-domain in intracellular transportation and compartmentalization of the toxin may be associated with the ability of the T-domain to form a proton channels and its ability to interact with COPI complex.http://ukrbiochemjournal.org/wp-content/uploads/2015/06/Labyntsev_3_14.pdfconfocal microscopydiphtheria toxinendocytosisfluorescent proteinsintracellular transportT-domain of diphtheria toxin |
| spellingShingle | А. J. Labyntsev D. V. Kolybo E. S. Yurchenko A. A. Kaberniuk N. V. Korotkevych S. V. Komisarenko Effect of the T-domain on intracellular transport of diphtheria toxin The Ukrainian Biochemical Journal confocal microscopy diphtheria toxin endocytosis fluorescent proteins intracellular transport T-domain of diphtheria toxin |
| title | Effect of the T-domain on intracellular transport of diphtheria toxin |
| title_full | Effect of the T-domain on intracellular transport of diphtheria toxin |
| title_fullStr | Effect of the T-domain on intracellular transport of diphtheria toxin |
| title_full_unstemmed | Effect of the T-domain on intracellular transport of diphtheria toxin |
| title_short | Effect of the T-domain on intracellular transport of diphtheria toxin |
| title_sort | effect of the t domain on intracellular transport of diphtheria toxin |
| topic | confocal microscopy diphtheria toxin endocytosis fluorescent proteins intracellular transport T-domain of diphtheria toxin |
| url | http://ukrbiochemjournal.org/wp-content/uploads/2015/06/Labyntsev_3_14.pdf |
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