Interaction of the COOH-terminal domain of the neurotensin receptor with a G protein does not control the phospholipase C activation but is involved in the agonist-induced internalization
Hermans, E.; Octave, J.N.; Maloteaux, J.M.
Molecular Pharmacology 49(2): 365-372
1996
ISSN/ISBN: 0026-895X
PMID: 8632771
Accession: 008893117
The agonist-induced internalization of the neurotensin receptor was studied in transfected Chinese hamster ovary cells expressing either the wild-type or a truncated rat neurotensin receptor, lacking the complete intracellular COOH-terminal end. Incubation of cells expressing the wild-type neurotensin receptor in the presence of the peptide resulted in a dramatic decrease in the (3H)neurotensin binding at the cell surface. This disappearance of cell surface binding sites resulted from the internalization of the receptor after the binding of the peptide. The receptor/peptide complexes were internalized in an intracellular compartment resistant to acid washes. The truncated receptor displayed high affinity binding properties for neurotensin in cell homogenates and activated phospholipase C as did the wild-type receptor. However, in cells expressing the truncated receptor, incubation with neurotensin only induced a partial decrease in cell surface binding, and internalization of the bound peptide was also impaired. On cell homogenates, the GTP analogue Gpp(NH)p was found to decrease the affinity of (3H)neurotensin for the wild-type receptor, whereas no similar effect was observed with the truncated receptor. These results show that the intracellular COOH-terminal region of the rat neurotensin receptor is not required for its functional coupling with intracellular G protein but is involved in the shift of the affinity of the receptor for the agonist, which occurs as a consequence of receptor activation and coupling. Because the truncated receptor was shown to internalize poorly, it may be proposed that internalization is not directly related to the activation of G protein but rather is a consequence of modification of receptor affinity, after activation by the agonist.