Linked Life Data

12071706

Source:http://linkedlifedata.com/resource/pubmed/id/12071706

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2002-6-19
pubmed:abstractText
The lutropin receptor consists of an extracellular N-terminal half and a membrane-associated C-terminal half. hCG initially binds the exodomain with a high affinity and the resulting complex is thought to interact with the endodomain through a secondary contact generating a hormonal signal. Therefore, the exodomain and endodomain are likely to associate directly or indirectly with each other, but lack of fruitful materials and technology has hampered knowledge about their physical relationship and contact sites. In this work, we engineered a double-recombinant (separate exodomain and endodomain) baculovirus system successfully expressing on the surface of insect cells high levels of split LH receptor, binding the hormone with high affinity and inducing cAMP synthesis. In contrast, the exodomain and endodomain expressed separately were mostly trapped in cells. Our data indicate that the exodomain and endodomain are disulfide linked in the split receptor. When the disulfide links were reduced, the split receptor still induced cAMP up to 60%, which raises the intriguing possibility of a residual induction activity of the endodomain in the absence of high-affinity ligand binding. Our results also underscore that the targeting and transport of the LH receptor to plasma membrane require both domains, whereas each domain is independently sufficient for folding. The expression level of functional lutropin receptors is the highest ever reported. Our system may also be useful for future studies requiring a high amount of soluble secreted exodomain.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
1046-5928
pubmed:author
pubmed:copyrightInfo
Copyright 2002 Elsevier Science (USA).
pubmed:issnType
Print
pubmed:volume
25
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
114-23
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:12071706-Animals, pubmed-meshheading:12071706-Cell Line, pubmed-meshheading:12071706-Cell Membrane, pubmed-meshheading:12071706-Cyclic AMP, pubmed-meshheading:12071706-DNA, Complementary, pubmed-meshheading:12071706-Disulfides, pubmed-meshheading:12071706-Dithiothreitol, pubmed-meshheading:12071706-Dose-Response Relationship, Drug, pubmed-meshheading:12071706-Immunoblotting, pubmed-meshheading:12071706-Insects, pubmed-meshheading:12071706-Ions, pubmed-meshheading:12071706-Kinetics, pubmed-meshheading:12071706-Ligands, pubmed-meshheading:12071706-Protein Binding, pubmed-meshheading:12071706-Protein Structure, Tertiary, pubmed-meshheading:12071706-Receptors, LH, pubmed-meshheading:12071706-Recombinant Proteins, pubmed-meshheading:12071706-Sodium Chloride, pubmed-meshheading:12071706-Swine, pubmed-meshheading:12071706-Time Factors
pubmed:year
2002
pubmed:articleTitle
Rescue of intracellularly trapped lutropin receptor exodomain by endodomain and reconstitution of a functional membrane receptor: interaction between exo- and endodomains.
pubmed:affiliation
Laboratoire de Physiologie des Cellules Cardiaques et Vasculaires, CNRS-UMR 6542, Tours, France.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't