dc.identifier.uri |
http://dx.doi.org/10.15488/191 |
|
dc.identifier.uri |
http://www.repo.uni-hannover.de/handle/123456789/213 |
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dc.contributor.author |
Breidert, Stephanie
|
|
dc.contributor.author |
Jacob, Ralf
|
|
dc.contributor.author |
Ngezahayo, Anaclet
|
|
dc.contributor.author |
Kolb, Hans-Albert
|
|
dc.contributor.author |
Naim, Hassan Y.
|
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dc.date.accessioned |
2016-02-02T12:03:12Z |
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dc.date.available |
2016-02-02T12:03:12Z |
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dc.date.issued |
2005-06-01 |
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dc.identifier.citation |
Breidert, S.; Jacob, R.; Ngezahayo, A.; Kolb, H.A.; Naim, H.Y.: Trafficking pathways of Cx49-GFP in living mammalian cells. In: Biological Chemistry 386 (2005), Nr. 2, S. 155-160. DOI: http://dx.doi.org/10.1515/BC.2005.019 |
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dc.description.abstract |
In the present study we examined the trafficking pathways of connexin49 (Cx49) fused to green fluorescent protein (GFP) in polar and non-polar cell lines. The Cx49 gene was isolated from ovine lens by RT-PCR. Cx49 cDNA was fused to GFP and the hybrid cDNA was transfected into several cell lines. After transfection of Cx49-GFP cDNA into HeLa cells, it was shown using the double whole-cell patch-clamp technique that the expressed fusion protein was still able to form conducting gap junction channels. Synthesis, assembly, and turnover of the Cx49-GFP hybrid protein were investigated using a pulse-chase protocol. A major 78-kDa protein band corresponding to Cx49-GFP could be detected with a turnover of 16-20 h and a half-life time of 10 h. The trafficking pathways of Cx49-GFP were monitored by confocal laser microscopy. Fusion proteins were localized in subcellular compartments, including the endoplasmic reticulum (ER), the ER-Golgi intermediate compartment, the Golgi apparatus, and the trans-Golgi network, as well as vesicles traveling towards the plasma membrane. Time-dependent sequential localization of Cx49-GFP in the ER and then the Golgi apparatus supports the notion of a slow turnover of Cx49-GFP compared to other connexins analyzed so far. Gap junction plaques resembling the usual punctuate distribution pattern could be demonstrated for COS-1 and MDCK cells. Basolateral distribution of Cx49-GFP was observed in polar MDCK cells, indicating specific sorting behavior of Cx49 in polarized cells. Together, this report describes the first characterization of biosynthesis and trafficking of lens Cx49. |
eng |
dc.description.sponsorship |
Fritz Thyssen-Stiftung |
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dc.language.iso |
eng |
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dc.publisher |
Berlin : Walter de Gruyter |
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dc.relation.ispartofseries |
Biological Chemistry 386 (2005), Nr. 2 |
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dc.rights |
Es gilt deutsches Urheberrecht. Das Dokument darf zum eigenen Gebrauch kostenfrei genutzt, aber nicht im Internet bereitgestellt oder an Außenstehende weitergegeben werden. Dieser Beitrag ist aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich. |
|
dc.subject |
connexin49 |
eng |
dc.subject |
green fluorescent protein |
eng |
dc.subject |
mammalian cells |
eng |
dc.subject |
protein assembly |
eng |
dc.subject |
protein transport |
eng |
dc.subject |
pulse-chase experiments |
eng |
dc.subject |
gap-junction channels |
eng |
dc.subject |
green fluorescent protein |
eng |
dc.subject |
hela-cells |
eng |
dc.subject |
molecular-cloning |
eng |
dc.subject |
connexin |
eng |
dc.subject |
lens |
eng |
dc.subject |
phosphorylation |
eng |
dc.subject |
permeability |
eng |
dc.subject |
degradation |
eng |
dc.subject |
cortex |
eng |
dc.subject.ddc |
570 | Biowissenschaften, Biologie
|
ger |
dc.title |
Trafficking pathways of Cx49-GFP in living mammalian cells |
eng |
dc.type |
Article |
|
dc.type |
Text |
|
dc.relation.essn |
1437-4315 |
|
dc.relation.issn |
1431-6730 |
|
dc.relation.doi |
http://dx.doi.org/10.1515/BC.2005.019 |
|
dc.bibliographicCitation.issue |
2 |
|
dc.bibliographicCitation.volume |
386 |
|
dc.bibliographicCitation.firstPage |
155 |
|
dc.bibliographicCitation.lastPage |
160 |
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dc.description.version |
publishedVersion |
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tib.accessRights |
frei zug�nglich |
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