The 100-kDa neurotensin receptor is gp95/sortilin, a non-G-protein-coupled receptor

J Biol Chem. 1998 Oct 9;273(41):26273-6. doi: 10.1074/jbc.273.41.26273.

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 (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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Vesicular Transport
  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Chromatography, Affinity
  • Cloning, Molecular
  • 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*

Substances

  • Adaptor Proteins, Vesicular Transport
  • DNA, Complementary
  • Membrane Glycoproteins
  • Nerve Tissue Proteins
  • Receptors, Cell Surface
  • GTP-Binding Proteins
  • sortilin