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- Title
A Cell-Free Translation and Proteoliposome Reconstitution System for Functional Analysis of Plant Solute Transporters.
- Authors
Akira Nozawa; Hideaki Nanamiya; Takuji Miyata; Nicole Linka; Yaeta Endo; Andreas P. M. Weber; Yuzuru Tozawa
- 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.
- Subjects
WHEAT; ARABIDOPSIS; PLANT cells &; tissues; GERM cells
- Publication
Plant & Cell Physiology, 2007, Vol 48, Issue 12, p1815
- ISSN
0032-0781
- Publication type
Article
- DOI
10.1093/pcp/pcm150