The 100-kDa Neurotensin Receptor Is gp95/Sortilin, A Non-G-Protein-coupled Receptor*
- Jean Mazella‡§,
- Nicole Zsürger‡,
- Valérie Navarro‡,
- Joëlle Chabry‡,
- Mourad Kaghad¶,
- Daniel Caput¶,
- Pascual Ferrara¶,
- Natalio Vita¶,
- Danielle Gully¶,
- Jean-Pierre Maffrand¶ and
- Jean-Pierre Vincent‡
- From the ‡Institut de Pharmacologie Moléculaire et Cellulaire, CNRS, UPR 0411, 660 route des Lucioles, Sophia Antipolis, 06560 Valbonne, France and ¶Sanofi Recherche, BP 137, 31676 Labège Cedex, France
Abstract
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 (K d = 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.
Footnotes
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↵* This work was supported by the Association pour la Recherche sur le Cancer.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵§ To whom correspondence should be addressed. Tel.: 33-4-93-95-77-61; Fax: 33-4-93-95-77-08; E-mail: mazella{at}ipmc.cnrs.fr.
- Abbreviations:
- NT
-
neurotensin
- CHAPS
-
3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid
- CHS
-
cholesteryl hemisuccinate
- PAGE
-
polyacrylamide gel electrophoresis
- DSS
-
disuccinimidyl suberate
- IGF-II
-
insulin-like growth factor
- Man-6-P
-
mannose 6-phosphate.
-
- Received July 8, 1998.
- Revision received August 4, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
