Source:http://linkedlifedata.com/resource/pubmed/id/17702749
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
41
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pubmed:dateCreated |
2007-10-8
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pubmed:abstractText |
The UT-A1 urea transporter mediates rapid transepithelial urea transport across the inner medullary collecting duct and plays a major role in the urinary concentrating mechanism. To transport urea, UT-A1 must be present in the plasma membrane. The purpose of this study was to screen for UT-A1-interacting proteins and to study the interactions of one of the identified potential binding partners with UT-A1. Using a yeast two-hybrid screen of a human kidney cDNA library with the UT-A1 intracellular loop (residues 409-594) as bait, we identified snapin, a ubiquitously expressed SNARE-associated protein, as a novel UT-A1 binding partner. Deletion analysis indicated that the C-terminal coiled-coil domain (H2) of snapin is required for UT-A1 interaction. Snapin binds to the intracellular loop of UT-A1 but not to the N- or C-terminal fragments. Glutathione S-transferase pulldown experiments and co-immunoprecipitation studies verified that snapin interacts with native UT-A1, SNAP23, and syntaxin-4 (t-SNARE partners), indicating that UT-A1 participates with the SNARE machinery in rat kidney inner medulla. Confocal microscopic analysis of immunofluorescent UT-A1 and snapin showed co-localization in both the cytoplasm and in the plasma membrane. When we co-injected UT-A1 with snapin cRNA in Xenopus oocytes, urea influx was significantly increased. In the absence of snapin, the influx was decreased when UT-A1 was combined with t-SNARE components syntaxin-4 and SNAP23. We conclude that UT-A1 may be linked to the SNARE machinery via snapin and that this interaction may be functionally and physiologically important for urea transport.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
12
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pubmed:volume |
282
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
30097-106
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pubmed:dateRevised |
2008-5-7
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pubmed:meshHeading |
pubmed-meshheading:17702749-Animals,
pubmed-meshheading:17702749-Biological Transport,
pubmed-meshheading:17702749-Cell Membrane,
pubmed-meshheading:17702749-DNA, Complementary,
pubmed-meshheading:17702749-Dogs,
pubmed-meshheading:17702749-Gene Library,
pubmed-meshheading:17702749-Humans,
pubmed-meshheading:17702749-Kidney,
pubmed-meshheading:17702749-Models, Biological,
pubmed-meshheading:17702749-Oocytes,
pubmed-meshheading:17702749-Protein Structure, Tertiary,
pubmed-meshheading:17702749-Rats,
pubmed-meshheading:17702749-Two-Hybrid System Techniques,
pubmed-meshheading:17702749-Vesicular Transport Proteins,
pubmed-meshheading:17702749-Xenopus
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pubmed:year |
2007
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pubmed:articleTitle |
The UT-A1 urea transporter interacts with snapin, a SNARE-associated protein.
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pubmed:affiliation |
Renal Division, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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