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Cloning and functional expression of a tetrabenazine sensitive vesicular monoamine transporter from bovine chromaffin granules
Author(s) -
Howell Mike,
Shirvan Anat,
Stern-Bach Yael,
Steiner-Mordoch Sonia,
Strasser Jane E.,
Dean Gary E.,
Schuldiner Shimon
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)80108-8
Subject(s) - tetrabenazine , vesicular monoamine transporter , vesicular monoamine transporter 2 , monoamine neurotransmitter , cloning (programming) , chemistry , vesicular transport protein , microbiology and biotechnology , transporter , biology , pharmacology , biochemistry , dopamine , neuroscience , serotonin , membrane , computer science , gene , vesicle , receptor , programming language
Using oligonucleotide primers derived from the vesicular monoamine transporters sequences, a cDNA predicted to encode the bovine chromaffin granule amine transporter has been cloned (b‐VMAT2). Surprisingly, its structure is more similar to the rat brain transporter (VMAT2), than to the rat adrenal counterpart (VMAT1). Unlike rat VMAT1, bovine VMAT2 appears to be expressed both in the adrenal medulla and the brain, as judged by Northern analysis. After modification/deletion of the seven amino acids at the N‐terminus of the protein it was expressed in a functional form. The order of affinity of the bovine VMAT2 transporter to substrates is: serotonin>dopamine = norepinephrine>epinephrine. Also, the recombinant bovine adrenal transporter is highly sensitive to tetrabenazine, in sharp contrast to the rat adrenal transporter. The findings indicate, therefore, a clear species variation in which structure and function of the bovine adrenal transporter resemble the rat brain protein, while its tissue distribution is distinct from both types of rat proteins. In addition, the predicted protein sequence is identical to the experimentally determined N‐terminus sequence of the purified vesicular amine transporter [Stern‐Bach et al. (1992) Proc. Natl. Acad. Sci. USA 89, 9730‐9733].