Chimeric rat/human neurotensin receptors localize a region of the receptor sensitive to binding of a novel, species-specific, picomolar affinity peptide

Bernadette Cusack, Karen Groshan, Daniel J Mc Cormick, Yuan-Ping Pang, Robin Perry, Chan Tran Phung, Terrance Souder, Elliott Richelsont

Research output: Contribution to journalArticle

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Abstract

Recently, we reported the development of a species-specific neurotensin analog that displays selective binding affinity at the rat and human neurotensin (NT) receptor, L-[3,2'-Nal11]NT(8-13) (where Nal is naphthylalanine) (NT19). We have developed another neurotensin analog, L- [3,1'-Nal11]NT(8-13), (NT34), that exhibits a 126-fold difference in binding affinities between the rat and human receptors. This compound differs from our previous reported species-specific ligand in the steric positioning of the naphthyl ring on the L-alanine side chain. For NT34, the observed K(d) values at the rat and human neurotensin receptors were 0.046 and 5.8 nM, respectively. In stimulating phosphatidylinositol turnover, the observed EC50 values were 2.8 nM and 130 nM in rat and human, respectively. We constructed a series of chimeric rat/human neurotensin receptor genes and expressed them by transient transfection into human embryonic kidney (HEK- 293) cells. Radioligand binding assays were then performed using neurotensin and NT34. Our results led us to propose a region of the neurotensin receptor that may be involved in determining species specificity, i.e. the transmembrane VI, the third extracellular loop, and transmembrane VII regions of the neurotensin receptor.

Original languageEnglish (US)
Pages (from-to)15054-15059
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number25
DOIs
StatePublished - 1996

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Neurotensin Receptors
neurotensin (8-13)
Rats
Neurotensin
Peptides
Species Specificity
Radioligand Assay
HEK293 Cells
Phosphatidylinositols
Alanine
Transfection
Assays
Genes
Ligands
Kidney

ASJC Scopus subject areas

  • Biochemistry

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Chimeric rat/human neurotensin receptors localize a region of the receptor sensitive to binding of a novel, species-specific, picomolar affinity peptide. / Cusack, Bernadette; Groshan, Karen; Mc Cormick, Daniel J; Pang, Yuan-Ping; Perry, Robin; Phung, Chan Tran; Souder, Terrance; Richelsont, Elliott.

In: Journal of Biological Chemistry, Vol. 271, No. 25, 1996, p. 15054-15059.

Research output: Contribution to journalArticle

Cusack, Bernadette ; Groshan, Karen ; Mc Cormick, Daniel J ; Pang, Yuan-Ping ; Perry, Robin ; Phung, Chan Tran ; Souder, Terrance ; Richelsont, Elliott. / Chimeric rat/human neurotensin receptors localize a region of the receptor sensitive to binding of a novel, species-specific, picomolar affinity peptide. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 25. pp. 15054-15059.
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