PMID: 11342177May 9, 2001Paper

Vesicular monoamine transporters heterologously expressed in the yeast Saccharomyces cerevisiae display high-affinity tetrabenazine binding

Biochimica Et Biophysica Acta
R Yelin, S Schuldiner

Abstract

A mammalian vesicular neurotransmitter transporter has been expressed in the yeast Saccharomyces cerevisiae. The gene encoding the rat vesicular monoamine transporter (rVMAT(1)) was cloned in several expression plasmids. The transporter was expressed at detectable levels only when short sequences using codons favored by S. cerevisiae were fused preceding the start of translation of rVMAT(1). The scarce expression of the wild-type protein was, most likely, due to the fact that part of the N-terminus of the protein is encoded by codons not preferred in S. cerevisiae. Furthermore, low growth temperatures increased rVMAT(1) expression and altered its processing. Whereas at 30 degrees C the protein is not glycosylated, at lower temperatures ( approximately 16 degrees C) half of the expressed transporters undergo core glycosylation. In addition, under these conditions the levels of protein expression significantly increase. Using a functional chimeric protein composed by VMAT and the green fluorescent protein (GFP), it is shown that the punctate pattern of intracellular distribution remains invariable at the different temperatures. Using a similar fusion sequence, the bovine VMAT isoform 2 (bVMAT(2)) was also expressed in yeast. The ...Continue Reading

References

Nov 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·J D EricksonB J Hoffman
Jan 1, 1991·Methods in Enzymology·M SchenaK R Yamamoto
Nov 17, 1989·Cell·R BallesterM Wigler
Aug 1, 1984·Molecular and Cellular Biology·M Johnston, R W Davis
Jul 1, 1984·Cell·I A WilsonR A Lerner
Jun 5, 1983·Journal of Molecular Biology·D Hanahan
Jan 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·D SchermanJ P Henry
Oct 27, 1995·The Journal of Biological Chemistry·A MerickelR H Edwards
Apr 1, 1995·Physiological Reviews·S SchuldinerM Linial
Feb 23, 1995·Nature·R HeimR Y Tsien
Aug 2, 1994·Molecular & General Genetics : MGG·A E Ehrenhofer-MurrayC Sengstag
Jan 1, 1995·Cold Spring Harbor Symposia on Quantitative Biology·D N HebertA Helenius
Jan 10, 1997·The Journal of Biological Chemistry·T W Loo, D M Clarke
Nov 18, 1998·Current Opinion in Structural Biology·E S Trombetta, A Helenius
Jun 11, 1999·The Journal of Biological Chemistry·C G TateM J Betenbaugh

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Citations

Apr 30, 2003·Journal of Structural Biology·Patrick J Loll
Dec 17, 2009·The Journal of Biological Chemistry·Yael Gros, Shimon Schuldiner
Jun 23, 2010·Protein Expression and Purification·Yael ElbazShimon Schuldiner
Apr 27, 2005·Biochemical and Biophysical Research Communications·Didier F PisaniIsabelle Mus-Veteau
Dec 6, 2005·Biochimica Et Biophysica Acta·Suphansa SawamiphakChuenchit Boonchird
May 26, 2009·Biochimica Et Biophysica Acta·Olivier JoubertIsabelle Mus-Veteau
Feb 22, 2008·American Journal of Physiology. Cell Physiology·Yoav AdamShimon Schuldiner
Sep 28, 2002·The Journal of Biological Chemistry·Hanbang ZhangPaul D Roepe
Jun 28, 2005·Brain Research. Molecular Brain Research·Amy K JassenBertha K Madras

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