Mazella J, Zsürger N, Navarro V, Chabry J, Kaghad M, Caput D, Ferrara P, Vita N, Gully D, Maffrand J P, Vincent J P
Institut de Pharmacologie Moléculaire et Cellulaire, CNRS, UPR 0411, 660 route des Lucioles, Sophia Antipolis, 06560 Valbonne, France.
J Biol Chem. 1998 Oct 9;273(41):26273-6. 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.
在本研究中，先前通过亲和层析从人脑中纯化得到的100 kDa神经降压素（NT）受体（Zsürger, N., Mazella, J., and Vincent, J. P. (1994) Brain Res. 639, 245 - 252），是从人脑海马cDNA文库中克隆得到的。该cDNA编码一个由833个氨基酸组成的蛋白质，与最近克隆的gp95/分拣蛋白100%相同，随后被命名为NT3受体-gp95/分拣蛋白。纯化蛋白的N末端与位于弗林蛋白酶切割位点之后的纯化gp95/分拣蛋白的序列相同。用克隆的cDNA转染的COS-7细胞经3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸增溶提取物，碘化NT与之结合具有饱和性且可逆，亲和力为10 - 15 nM。NT3受体-gp95/分拣蛋白在细胞内囊泡中的定位与先前用纯化受体和gp95/分拣蛋白得到的结果一致。亲和标记和结合实验表明，110 kDa的NT3受体通过与弗林蛋白酶共转染可部分转化为亲和力更高（Kd = 0.3 nM）的100 kDa蛋白受体。这种100 kDa的NT受体对应于受体的成熟形式。NT3/gp95/分拣蛋白是首个不属于G蛋白偶联受体超家族的跨膜神经肽受体。
