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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
44
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pubmed:dateCreated |
1996-12-26
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pubmed:abstractText |
To obtain data of the bradykinin B2 receptor's agonist binding site, we used a combined approach of affinity labeling and "immunoidentification" of receptor fragments generated by cyanogen bromide cleavage. Domain-specific antibodies to the various extracellular receptor domains were applied to detect receptor fragments with covalently attached [125I-Tyr8]bradykinin. As a cross-linker we used the homobifunctional reagent disuccinimidyl tartarate (DST), which reacts preferentially with primary amines. With this technique a [125I-Tyr8]bradykinin-labeled receptor fragment derived from the third extracellular domain was identified. The epsilon-amino group of lysine (Lys172) of the human B2 receptor provides the only primary amino group within this receptor fragment. This strongly suggests that DST attached the N-terminal amino group of [Tyr8]bradykinin to Lys172 of the human B2 receptor. Next we asked whether DST attaches [Tyr8]bradykinin to the analogous residue, Lys174 of the rat B2 receptor, which is 81% identical to the human B2 receptor, and we attempted to label the wild-type rat B2 receptor and a rat B2 receptor mutant where Lys174 had been exchanged for alanine. Affinity labeling of the wild-type rat B2 receptor worked efficiently, whereas DST did not attach detectable amounts of [125I-Tyr8]bradykinin to the K174A rat B2 receptor mutant. Taken together these observations indicate that the N-terminal amino group of [Tyr8]bradykinin is bound to analogous positions of the rat and of the human B2 receptor, i.e. [Tyr8]bradykinin's N terminus is bound adjacent to Lys172 of the human and Lys174 of the rat B2 receptor.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Affinity Labels,
http://linkedlifedata.com/resource/pubmed/chemical/Bradykinin,
http://linkedlifedata.com/resource/pubmed/chemical/Cross-Linking Reagents,
http://linkedlifedata.com/resource/pubmed/chemical/Iodine Radioisotopes,
http://linkedlifedata.com/resource/pubmed/chemical/Lysine,
http://linkedlifedata.com/resource/pubmed/chemical/Receptor, Bradykinin B2,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Bradykinin,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
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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 |
1
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pubmed:volume |
271
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
27382-7
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:8910316-Affinity Labels,
pubmed-meshheading:8910316-Amino Acid Sequence,
pubmed-meshheading:8910316-Animals,
pubmed-meshheading:8910316-Binding Sites,
pubmed-meshheading:8910316-Bradykinin,
pubmed-meshheading:8910316-COS Cells,
pubmed-meshheading:8910316-Cell Line,
pubmed-meshheading:8910316-Cross-Linking Reagents,
pubmed-meshheading:8910316-Humans,
pubmed-meshheading:8910316-Iodine Radioisotopes,
pubmed-meshheading:8910316-Lysine,
pubmed-meshheading:8910316-Molecular Sequence Data,
pubmed-meshheading:8910316-Protein Structure, Secondary,
pubmed-meshheading:8910316-Rats,
pubmed-meshheading:8910316-Receptor, Bradykinin B2,
pubmed-meshheading:8910316-Receptors, Bradykinin,
pubmed-meshheading:8910316-Recombinant Proteins,
pubmed-meshheading:8910316-Transfection
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pubmed:year |
1996
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pubmed:articleTitle |
The N-terminal amino group of [Tyr8]bradykinin is bound adjacent to analogous amino acids of the human and rat B2 receptor.
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pubmed:affiliation |
Institute of Physiological Chemistry and Pathobiochemistry, University of Mainz, 55099 Germany.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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