Chicken liver contains a large quantity of a G-protein-linked neurotensin receptor.
Using 125I-labeled neurotensin (NT), chicken liver was found to contain high affinity, G-protein-linked receptors directed specifically towards the bioactive C-terminal portion of NT. Binding was proportional to membrane and optimal at pH 7.5. The apparent Kd (approximately 91 pM) for this single class of binding sites was similar to Kds reported for the high-affinity components of NT binding to mammalian brain and intestinal membranes. However, the binding capacity (Bmax, approximately 2.3 pmol/mg) was 10-100 times higher than values reported for these mammalian tissues. Binding was inhibited by GTP analogues and by treatment with pertussis toxin but not by cholera toxin. Treatments with alkaline solutions, shown to inactivate G-proteins, decreased subsequent binding at pH 7.5. Whereas low concentrations of Mg2+ (optimum, approximately 0.5 mM) enhanced NT binding, concentrations of 5 mM and above were inhibitory. Cross-linking of 125I-labeled NT to liver membranes using glutaraldehyde specifically labeled two substances of approximately 52 and approximately 90 kDa, which could represent different binding proteins or complexes. These data demonstrate the presence in chicken liver of large amounts of high-affinity NT receptor(s) coupled to pertussis toxin-sensitive G-protein(s).
Mitra
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Physiology Department, University of Massachusetts Medical Center, Worcester 01655, USA.
Carraway
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D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002645:Chickens / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000737:chemistry*; D007457:Iodine Radioisotopes; D016014:Linear Models; D008099:Liver / Q000737:chemistry*; D008566:Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D009496:Neurotensin / Q000378:metabolism*; D018028:Receptors, Neurotensin / Q000032:analysis* / Q000737:chemistry
D007457:Iodine Radioisotopes; D018028:Receptors, Neurotensin; D009496:Neurotensin; D019204:GTP-Binding Proteins