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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
34
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pubmed:dateCreated |
2005-8-22
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pubmed:abstractText |
Gap junction channels play an important role in cell growth control, secretion and embryonic development. Gap junctional communication and channel assembly can be regulated by protein-protein interaction with kinases and phosphatases. We have utilized tandem mass spectrometry (MS/MS) sequence analysis as a screen to identify proteins from cell lysates that interact with the C-terminal cytoplasmic region of connexin 43 (Cx43). MS/MS analysis of tryptic fragments yielded several proteins including zona occludens-1 (ZO-1), a structural protein previously identified to interact with Cx43, and ZO-2, a potential novel interacting partner. We confirmed the interaction of ZO-2 with Cx43 by using a combination of fusion protein "pull down," co-immunoprecipitation, and co-localization experiments. We show that the C-terminal region of Cx43 is necessary for interaction with the PDZ2 domain of ZO-2. Far Western analysis revealed that ZO-2 can directly bind to Cx43 independent of other interacting partners. Immunofluorescence studies indicate that both ZO-1 and ZO-2 can co-localize with Cx43 within the plasma membrane at apparent gap junctional structures. We examined Cx43 interaction with ZO-1 and ZO-2 at different stages of the cell cycle and found that Cx43 had a strong preference for interaction with ZO-1 during G0, whereas ZO-2 interaction occurred approximately equally during G0 and S phases. Since essentially all of the Cx43 in G0 cells is assembled into Triton X-100-resistant junctions, Cx43-ZO-1 interaction may contribute to their stability.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0021-9258
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
26
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pubmed:volume |
280
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
30416-21
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:15980428-Animals,
pubmed-meshheading:15980428-Blotting, Western,
pubmed-meshheading:15980428-Cell Cycle,
pubmed-meshheading:15980428-Cell Line,
pubmed-meshheading:15980428-Cell Membrane,
pubmed-meshheading:15980428-Chromatography,
pubmed-meshheading:15980428-Connexin 43,
pubmed-meshheading:15980428-Cytoplasm,
pubmed-meshheading:15980428-Detergents,
pubmed-meshheading:15980428-Epithelial Cells,
pubmed-meshheading:15980428-Gap Junctions,
pubmed-meshheading:15980428-Glutathione Transferase,
pubmed-meshheading:15980428-Immunoblotting,
pubmed-meshheading:15980428-Immunoprecipitation,
pubmed-meshheading:15980428-Kidney,
pubmed-meshheading:15980428-Mass Spectrometry,
pubmed-meshheading:15980428-Membrane Proteins,
pubmed-meshheading:15980428-Microscopy, Confocal,
pubmed-meshheading:15980428-Microscopy, Fluorescence,
pubmed-meshheading:15980428-Octoxynol,
pubmed-meshheading:15980428-Phosphoproteins,
pubmed-meshheading:15980428-Plasmids,
pubmed-meshheading:15980428-Protein Binding,
pubmed-meshheading:15980428-Protein Structure, Tertiary,
pubmed-meshheading:15980428-Rats,
pubmed-meshheading:15980428-Recombinant Fusion Proteins,
pubmed-meshheading:15980428-S Phase,
pubmed-meshheading:15980428-Trypsin
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pubmed:year |
2005
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pubmed:articleTitle |
Connexin 43 interacts with zona occludens-1 and -2 proteins in a cell cycle stage-specific manner.
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pubmed:affiliation |
Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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