PMID: 9191107Jun 1, 1997Paper

Identification in the rat neurotensin receptor of amino-acid residues critical for the binding of neurotensin

Brain Research. Molecular Brain Research
J M BottoJ Mazella

Abstract

In order to identify charged amino-acid residues of the cloned rat brain neurotensin (NT) receptor (NTR) that are critical for NT binding, we performed site-directed mutagenesis on the cDNA encoding this protein, followed by transient expression into mammalian COS-7 cells and in Xenopus laevis oocytes. Point substitutions of charged residues in the N-terminal part and in the 2nd and 3rd extracellular loop of the receptor either did not affect (125)I-Tyr3-NT binding or resulted in a decrease in binding affinity by a factor of 2-3. Mutations of amino acids Asp113 in the second transmembrane domain (TM) and of Arg149 or Asp150 in TM III yielded receptors that bound NT as efficiently as the native receptor. By contrast, replacement of the Asp139 residue in the 1st extracellular loop, or of Arg143 or Arg327-Arg328 residues at the top of TM III and in TM VI, respectively, completely abolished ligand binding. Confocal and EM immunocytochemical studies of the expression of these affected receptors, tagged with the C-terminal sequence of the vesicular stomatitis virus glycoprotein (VSV-G), indicated that this loss of binding was not due to altered receptor expression or to their improper insertion into the plasma membrane. When these mu...Continue Reading

References

Jan 1, 1992·DNA and Cell Biology·W C ProbstS C Sealfon
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Jan 1, 1994·Annual Review of Biochemistry·C D StraderR A Dixon
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Citations

Jan 24, 2014·Proceedings of the National Academy of Sciences of the United States of America·Pascal EgloffAndreas Plückthun
Jun 26, 1998·Journal of Clinical Laser Medicine & Surgery
May 26, 2009·ACS Chemical Biology·Rebecca M MyersSteven V Ley
Jul 3, 1999·Trends in Pharmacological Sciences·J P VincentP Kitabgi
Aug 25, 2020·ACS Pharmacology & Translational Science·Fabian BumbakDaniel J Scott
Jan 5, 2011·General and Comparative Endocrinology·Makoto NumaoMinoru Tanaka

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