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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
3
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pubmed:dateCreated |
1998-11-23
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pubmed:abstractText |
A chimeric cDNA construct encoding the extracellular amino-terminal domain (ECD) of the thyrotropin receptor fused to the signal for addition of glycosylphosphatidylinositol from the Thy-1 gene directs efficient expression of the ECD at the plasma membrane of transfected CHO cells. A cell line (GT14) expressing over 106 receptors/cell was isolated, which allows direct detection, by flow cytometry, of autoantibodies from the majority of patients with Graves' disease or autoimmune idiopathic myxedema. Treatment of GT14 cells with a glycosylphosphatidylinositol-specific phospholipase C (PI-PLC) releases a soluble 80 kDa molecule which neutralizes the autoantibodies from Graves patients. Whereas it does not bind TSH when released from the cells by PI-PLC in free form, the soluble ECD displays clear TSH binding activity when it is released as a complex with a monoclonal antibody recognizing a conformational epitope of the ECD. Our results allow production of bioactive ECD of the thyrotropin receptor in high yield, with possible applications in structural analyses.
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Thy-1,
http://linkedlifedata.com/resource/pubmed/chemical/Glycosylphosphatidylinositols,
http://linkedlifedata.com/resource/pubmed/chemical/Immunoglobulins, Thyroid-Stimulating,
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphatidylinositol...,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoinositide Phospholipase C,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Thyrotropin,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Thyrotropin,
http://linkedlifedata.com/resource/pubmed/chemical/Type C Phospholipases
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0014-5793
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
9
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pubmed:volume |
436
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
427-33
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:9801163-Amino Acid Sequence,
pubmed-meshheading:9801163-Animals,
pubmed-meshheading:9801163-Antigens, Thy-1,
pubmed-meshheading:9801163-Autoimmune Diseases,
pubmed-meshheading:9801163-CHO Cells,
pubmed-meshheading:9801163-Cell Line,
pubmed-meshheading:9801163-Cell Membrane,
pubmed-meshheading:9801163-Cricetinae,
pubmed-meshheading:9801163-Glycosylphosphatidylinositols,
pubmed-meshheading:9801163-Graves Disease,
pubmed-meshheading:9801163-Humans,
pubmed-meshheading:9801163-Hypothyroidism,
pubmed-meshheading:9801163-Immunoglobulins, Thyroid-Stimulating,
pubmed-meshheading:9801163-Molecular Sequence Data,
pubmed-meshheading:9801163-Neutralization Tests,
pubmed-meshheading:9801163-Peptide Fragments,
pubmed-meshheading:9801163-Phosphatidylinositol Diacylglycerol-Lyase,
pubmed-meshheading:9801163-Phosphoinositide Phospholipase C,
pubmed-meshheading:9801163-Polymerase Chain Reaction,
pubmed-meshheading:9801163-Protein Conformation,
pubmed-meshheading:9801163-Receptors, Thyrotropin,
pubmed-meshheading:9801163-Recombinant Proteins,
pubmed-meshheading:9801163-Thyrotropin,
pubmed-meshheading:9801163-Transfection,
pubmed-meshheading:9801163-Type C Phospholipases
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pubmed:year |
1998
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pubmed:articleTitle |
Production of bioactive amino-terminal domain of the thyrotropin receptor via insertion in the plasma membrane by a glycosylphosphatidylinositol anchor.
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pubmed:affiliation |
I.R.I.B.H.N., Faculté de Médecine, University of Brussels, and Euroscreen s.a., Belgium.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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