The 100-kDa Neurotensin Receptor Is gp95/Sortilin, A Non-G-Protein-coupled Receptor

• Published in: The Journal of biological chemistry Vol. 273; no. 41; pp. 26273 - 26276
• Main Authors: Mazella, Jean, Zsürger, Nicole, Navarro, Valérie, Chabry, Joëlle, Kaghad, Mourad, Caput, Daniel, Ferrara, Pascual, Vita, Natalio, Gully, Danielle, Maffrand, Jean-Pierre, Vincent, Jean-Pierre
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 09.10.1998; American Society for Biochemistry and Molecular Biology
• Subjects: Adaptor Proteins, Vesicular Transport; Amino Acid Sequence; Animals; Chromatography, Affinity; Cloning, Molecular; COS Cells; DNA, Complementary; Electrophoresis, Polyacrylamide Gel; GTP-Binding Proteins - metabolism; Humans; Membrane Glycoproteins - chemistry; Membrane Glycoproteins - isolation & purification; Membrane Glycoproteins - metabolism; Molecular Sequence Data; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - isolation & purification; Nerve Tissue Proteins - metabolism; Protein Binding; Receptors, Cell Surface - chemistry; Receptors, Cell Surface - isolation & purification; Receptors, Cell Surface - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.41.26273

In this work, the 100-kDa neurotensin (NT) receptor previously purified from human brain by affinity chromatography (Zsürger, N., Mazella, J., and Vincent, J. P. (1994) Brain Res. 639, 245–252) was cloned from a human brain cDNA library. This cDNA encodes a 833-amino acid protein 100% identical to the recently cloned gp95/sortilin and was then designated NT3 receptor-gp95/sortilin. The N terminus of the purified protein is identical to the sequence of the purified gp95/sortilin located immediately after the furin cleavage site. The binding of iodinated NT to 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid-solubilized extracts of COS-7 cells transfected with the cloned cDNA was saturable and reversible with an affinity of 10–15 nm. The localization of the NT3 receptor-gp95/sortilin into intracellular vesicles was in agreement with previous results obtained with the purified receptor and with gp95/sortilin. Affinity labeling and binding experiments showed that the 110-kDa NT3 receptor can be partly transformed into a higher affinity (Kd = 0.3 nm) 100-kDa protein receptor by cotransfection with furin. This 100-kDa NT receptor corresponded to the mature form of the receptor. The NT3/gp95/sortilin protein is the first transmembrane neuropeptide receptor that does not belong to the superfamily of G-protein-coupled receptors.