Cloning and expression of the vesamicol binding protein from the marine ray Torpedo

FEBS Letters

Varoqui, Hélène³  Erickson, Jeffrey D.²  Usdin, Ted B.²  Eiden, Lee E.²  Diebler, Marie-Françoise³  Bonner, Tom I.²  Meunier, François-Marie³  Rand, James B.¹ 

[1] Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA;Laboratory of Cell Biology, NIMHINIH, Bethesda, MD 20892, USA;Département de Neurochimie, Laboratoire de Neurobiologie Cellulaire, CNRS, 91190 Gif sur Yvette, France

关键词: Vesicular acetylcholine transporter;    Vesamicol;    Torpedo electric lobe;    Caenorhabditis elegans UNC-17;   

DOI  :  10.1016/0014-5793(94)80592-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Complementary DNA clones corresponding to a messenger RNA encoding a 56 kDa polypeptide have been obtained from Torpedo marmorata and Torpedo ocellata electric lobe libraries, by homology screening with a probe obtained from the putative acetylcholine transporter from the nematode Caenorhabditis elegans. The Torpedo proteins display approximately 50% overall identity to the C. elegans unc-17 protein and 43% identity to the two vesicle monoamine transporters (VMAT1 and VMAT2). This family of proteins is highly conserved within 12 domains which potentially span the vesicle membrane, with little similarity within the putative intraluminal glycosylated loop and at the N- and C-termini. The ~ 3.0 kb mRNA species is specifically expressed in the brain and highly enriched in the electric lobe of Torpedo. The Torpedo protein, expressed in CV-1 fibroblast cells, possesses a high-affinity binding site for vesamicol (K _d = 6 nM), a drug which blocks in vitro and in vivo acetylcholine accumulation in cholinergic vesicles.

【 授权许可】

Unknown