Source:http://linkedlifedata.com/resource/pubmed/id/15319442
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| Predicate | Object |
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| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
45
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| pubmed:dateCreated |
2004-11-1
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| pubmed:abstractText |
To identify molecules that might contribute to V2 vasopressin receptor (V2R) trafficking or signaling, we searched for novel interacting proteins with this receptor. Preliminary data, using the V2R C terminus as bait in a yeast two-hybrid screen, revealed calmodulin as a binding partner. Because calmodulin interacts with other G protein-coupled receptors, we explored this interaction and its possible functional relevance in greater detail. A Ca2+ -dependent interaction occurs between calmodulin-linked agarose and the holo-V2R as well as the V2R C terminus. Truncation and site-directed mutagenesis of the V2R C terminus revealed an involvement of an RGR sequence in this interaction. NMR studies showed that a peptide fragment of the V2R C terminus containing the RGR sequence binds to calmodulin in a Ca2+ -dependent manner with a Kd < or =1.5 microm; concentration-dependent binding of the V2R C terminus to calmodulin-agarose was used to estimate a Kd value of approximately 200 nm for this entire C-terminal sequence as expressed in mammalian cells. Madin-Darby canine kidney II cells stably expressing either wild type or a mutant V2R, in which the RGR C-terminal sequence was mutated to alanines (AAA V2R), revealed that the steady-state localization and agonist-induced internalization of the AAA V2R resembled that of the wild type V2R in polarized Madin-Darby canine kidney II cells. V2R binding of agonist similarly was unchanged in the AAA V2R, as was the concentration response for arginine vasopressin (AVP)-stimulated cAMP accumulation. Most interestingly, AVP-induced increases in intracellular Ca2+ observed for the wild type V2R were virtually eliminated for the AAA V2R. Taken together, the data suggest that a C-terminal region of the V2R important for calmodulin interaction is also important in modulation of V2R elevation of intracellular Ca2+, a prerequisite for AVP-induced fusion of aquaporin-containing vesicles with the apical surface of renal epithelial cells.
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| pubmed:grant | |
| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Calcium,
http://linkedlifedata.com/resource/pubmed/chemical/Calmodulin,
http://linkedlifedata.com/resource/pubmed/chemical/Cyclic AMP,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/Detergents,
http://linkedlifedata.com/resource/pubmed/chemical/Glutathione Transferase,
http://linkedlifedata.com/resource/pubmed/chemical/Peptides,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Vasopressin,
http://linkedlifedata.com/resource/pubmed/chemical/Sepharose,
http://linkedlifedata.com/resource/pubmed/chemical/Vasopressins
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| pubmed:status |
MEDLINE
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| pubmed:month |
Nov
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| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
5
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| pubmed:volume |
279
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
46969-80
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| pubmed:dateRevised |
2007-11-14
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| pubmed:meshHeading |
pubmed-meshheading:15319442-Amino Acid Sequence,
pubmed-meshheading:15319442-Animals,
pubmed-meshheading:15319442-Binding, Competitive,
pubmed-meshheading:15319442-COS Cells,
pubmed-meshheading:15319442-Calcium,
pubmed-meshheading:15319442-Calmodulin,
pubmed-meshheading:15319442-Cell Line,
pubmed-meshheading:15319442-Cell Membrane,
pubmed-meshheading:15319442-Cyclic AMP,
pubmed-meshheading:15319442-DNA, Complementary,
pubmed-meshheading:15319442-Detergents,
pubmed-meshheading:15319442-Dogs,
pubmed-meshheading:15319442-Dose-Response Relationship, Drug,
pubmed-meshheading:15319442-Endocytosis,
pubmed-meshheading:15319442-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:15319442-Glutathione Transferase,
pubmed-meshheading:15319442-Humans,
pubmed-meshheading:15319442-Hydrogen-Ion Concentration,
pubmed-meshheading:15319442-Kinetics,
pubmed-meshheading:15319442-Magnetic Resonance Spectroscopy,
pubmed-meshheading:15319442-Molecular Sequence Data,
pubmed-meshheading:15319442-Peptides,
pubmed-meshheading:15319442-Protein Binding,
pubmed-meshheading:15319442-Protein Conformation,
pubmed-meshheading:15319442-Protein Structure, Tertiary,
pubmed-meshheading:15319442-Receptors, Vasopressin,
pubmed-meshheading:15319442-Sepharose,
pubmed-meshheading:15319442-Sequence Homology, Amino Acid,
pubmed-meshheading:15319442-Signal Transduction,
pubmed-meshheading:15319442-Time Factors,
pubmed-meshheading:15319442-Transfection,
pubmed-meshheading:15319442-Two-Hybrid System Techniques,
pubmed-meshheading:15319442-Vasopressins
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| pubmed:year |
2004
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| pubmed:articleTitle |
Calmodulin interacts with the V2 vasopressin receptor: elimination of binding to the C terminus also eliminates arginine vasopressin-stimulated elevation of intracellular calcium.
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| pubmed:affiliation |
Department of Pharmacology, Vanderbilt University Medical Center, Nashville, Tennessee 37232-6600, USA.
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| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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