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rdf:type |
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lifeskim:mentions |
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pubmed:dateCreated |
1996-1-25
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pubmed:abstractText |
We have utilized cell-free translation in rabbit-reticulocyte lysate supplemented with canine pancreatic microsomal membranes to study the processing and membrane topology of the rat ionotropic glutamate receptor subunit GluR1. In vitro-synthesized RNA encoding GluR1 was translated to yield a primary translation product with an apparent molecular mass of 99 kDa. In the presence of microsomal membranes this protein was processed to give a band of 107 kDa. Treatment with peptide-N-glycosidase F showed that this increase in molecular mass was due to N-linked glycosylation. Incubation of the processed receptor with proteinase K revealed the presence of a 68 kDa protease-resistant band which decreased to 56 kDa following deglycosylation. A deletion mutant (GluR1M1) lacking the predicted transmembrane domains was fully translocated across the microsomal membrane and protected from the action of the protease. The mutant and wild-type receptor could be immunoprecipitated by anti-peptide antibodies directed against the C-terminus. Following translocation of the wild-type and mutant receptor across the microsomal membrane and treatment with proteinase K the antibody binding to GluR1 was abolished, but was retained for GluR1M1. These data allow identification of the orientation of the N- and C-termini of GluR1 within the microsome; results which are consistent with an extracellular N-terminal and intracellular C-terminal localization following incorporation into the plasma membrane.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-1314044,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-1374769,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-1699275,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-2166337,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-2480522,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-2558391,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-2844410,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-3896128,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-4063349,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-6361452,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-6656655,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7518502,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7527641,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7539962,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7542368,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7838318,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7857646,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-7993626,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-8041762,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-8094892,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-8163463,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-8188697,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-8316301,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8526855-8492909
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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 |
Dec
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pubmed:issn |
0264-6021
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
312 ( Pt 2)
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
451-6
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:8526855-Animals,
pubmed-meshheading:8526855-Cell-Free System,
pubmed-meshheading:8526855-Cloning, Molecular,
pubmed-meshheading:8526855-Dogs,
pubmed-meshheading:8526855-Glycosylation,
pubmed-meshheading:8526855-Intracellular Membranes,
pubmed-meshheading:8526855-Macromolecular Substances,
pubmed-meshheading:8526855-Microsomes,
pubmed-meshheading:8526855-Models, Structural,
pubmed-meshheading:8526855-Molecular Weight,
pubmed-meshheading:8526855-Mutagenesis,
pubmed-meshheading:8526855-Protein Biosynthesis,
pubmed-meshheading:8526855-Protein Structure, Secondary,
pubmed-meshheading:8526855-Rabbits,
pubmed-meshheading:8526855-Rats,
pubmed-meshheading:8526855-Receptors, AMPA,
pubmed-meshheading:8526855-Recombinant Proteins,
pubmed-meshheading:8526855-Reticulocytes,
pubmed-meshheading:8526855-Sequence Deletion,
pubmed-meshheading:8526855-Transcription, Genetic
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pubmed:year |
1995
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pubmed:articleTitle |
An investigation of the membrane topology of the ionotropic glutamate receptor subunit GluR1 in a cell-free system.
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pubmed:affiliation |
Department of Pharmacology, Medical School, Edgbaston, Birmingham, U.K.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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