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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
4
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pubmed:dateCreated |
2003-9-1
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pubmed:abstractText |
A protein corresponding to the extracellular 1-209 domain of the alpha-subunit of the nicotine acetylcholine receptor from the electric organ of Torpedo californica was prepared using the corresponding cDNA domain by culturing Escherichia coli cells on a synthetic medium supplemented with 5-fluoro-L-tryptophan. The presence of a (His)6 fragment preceding the 1-209 sequence allowed purification of the protein isolated from inclusion bodies by affinity chromatography on Ni-NTA Agarose. The incorporation of 5-fluorotryptophan residues was found by 19F NMR to be approximately 50%. The spectrum of the protein reduced under denaturing conditions and subsequently reoxidized in a dilute solution under denaturing conditions in the presence of 0.05% SDS was sufficiently resolved, which allowed partial assignment of 19F resonances using the Trp60Phe mutant protein. The ability of the prepared domains to specifically bind snake alpha-neurotoxins was demonstrated with the use of radioiodinated alpha-bungarotoxin and trifluoroacetylated alpha-cobratoxin.
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pubmed:language |
rus
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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/5-fluorotryptophan,
http://linkedlifedata.com/resource/pubmed/chemical/Bungarotoxins,
http://linkedlifedata.com/resource/pubmed/chemical/Cobra Neurotoxin Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorine,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Subunits,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Cholinergic,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Sodium Dodecyl Sulfate,
http://linkedlifedata.com/resource/pubmed/chemical/Tryptophan,
http://linkedlifedata.com/resource/pubmed/chemical/alpha-cobratoxin
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pubmed:status |
MEDLINE
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pubmed:issn |
0132-3423
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
29
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
384-90
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:12947759-Animals,
pubmed-meshheading:12947759-Binding Sites,
pubmed-meshheading:12947759-Bungarotoxins,
pubmed-meshheading:12947759-Chromatography, Affinity,
pubmed-meshheading:12947759-Cobra Neurotoxin Proteins,
pubmed-meshheading:12947759-Escherichia coli,
pubmed-meshheading:12947759-Extracellular Matrix,
pubmed-meshheading:12947759-Fluorine,
pubmed-meshheading:12947759-Magnetic Resonance Spectroscopy,
pubmed-meshheading:12947759-Mutation,
pubmed-meshheading:12947759-Protein Denaturation,
pubmed-meshheading:12947759-Protein Folding,
pubmed-meshheading:12947759-Protein Structure, Tertiary,
pubmed-meshheading:12947759-Protein Subunits,
pubmed-meshheading:12947759-Receptors, Cholinergic,
pubmed-meshheading:12947759-Recombinant Proteins,
pubmed-meshheading:12947759-Sodium Dodecyl Sulfate,
pubmed-meshheading:12947759-Torpedo,
pubmed-meshheading:12947759-Tryptophan
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pubmed:articleTitle |
[5-fluoro-tryptophan-containing N-terminal domain of the alpha-subunit of the Torpedo californica acetylcholine receptor: preparation in E. coli and 19F NMR study].
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pubmed:affiliation |
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, GSP Moscow, 117997 Russia. t_alex@freemail.ru
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pubmed:publicationType |
Journal Article,
English Abstract
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