Skip Navigation


Plant and Cell Physiology Advance Access first published online on November 2, 2007
This version published online on November 5, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm150
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
48/12/1815    most recent
pcm150v2
pcm150v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Nozawa, A.
Right arrow Articles by Tozawa, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nozawa, A.
Right arrow Articles by Tozawa, Y.
Agricola
Right arrow Articles by Nozawa, A.
Right arrow Articles by Tozawa, Y.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

A cell-free translation and proteoliposome reconstitution system for functional analysis of plant solute transporters

Akira Nozawa1, Hideaki Nanamiya1, Takuji Miyata2, Nicole Linka3, Yaeta Endo1,2, Andreas P. M. Weber3 and Yuzuru Tozawa1,2

1Cell-Free Science and Technology Research Center, and The Venture Business Laboratory, Ehime University, Matsuyama 790-8577, Japan
2Department of Applied Chemistry, Faculty of Engineering, Ehime University, Matsuyama 790-8577, Japan
3Institute for Plant Biochemistry, Heinrich-Heine-University, Geb. 26.03.01, Universitätsstraße 1, D-40225 Düsseldorf, Germany

Corresponding author: Yuzuru Tozawa, Division of Biomolecular Engineering, Cell-Free Science and Technology Research Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime 790-8577, Japan. Tel.: +81-89-927-8274. Fax: +81-89-927-8276. E-mail: tozaway{at}ccr.ehime-u.ac.jp


  Abstract

We describe here a novel proteoliposome reconstitution system for functional analysis of plant membrane transporters that is based on a modified wheat germ cell-free translation system. We established optimized conditions for the reconstitution system with Arabidopsis thaliana phosphoenolpyruvate/phosphate translocator 1 (AtPPT1) as a model transporter. A high activity of AtPPT1 was achieved by synthesis of the protein in the presence of both a detergent such as Brij35 and liposomes. We also determined the substrate specificities of three putative rice PPT homologs with this system. The cell-free proteoliposome reconstitution system provides a valuable tool for functional analysis of transporter proteins.

Keywords: cell-free translation system - phosphoenolpyruvate - proteoliposome - transporter - wheat germ

(Received September 26, 2007; Accepted October 29, 2007)
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.