Trafficking pathways of Cx49-GFP in living mammalian cells

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dc.identifier.uri Breidert, Stephanie Jacob, Ralf Ngezahayo, Anaclet Kolb, Hans-Albert Naim, Hassan Y. 2016-02-02T12:03:12Z 2016-02-02T12:03:12Z 2005-06-01
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:
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
dc.language.iso eng
dc.publisher Berlin : Walter de Gruyter
dc.relation.ispartofseries Biological Chemistry 386 (2005), Nr. 2
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.bibliographicCitation.issue 2
dc.bibliographicCitation.volume 386
dc.bibliographicCitation.firstPage 155
dc.bibliographicCitation.lastPage 160
dc.description.version publishedVersion
tib.accessRights frei zug�nglich

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