<i>In Vitro</i>Protein Import of a Putative Amino Acid Transporter from<i>Arabidopsis thaliana</i>into Chloroplasts and Its Suborganellar Localization

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  • SATTASUK Kwanchanok
    The United Graduate School of Agricultural Sciences, Ehime University
  • NOZAWA Akira
    Cell-Free Science and Technology Research Center, Ehime University Satellite Business Venture, Ehime University
  • TOZAWA Yuzuru
    Cell-Free Science and Technology Research Center, Ehime University Satellite Business Venture, Ehime University
  • KAKINUMA Yoshimi
    The United Graduate School of Agricultural Sciences, Ehime University Faculty of Agriculture, Ehime University
  • AKITA Mitsuru
    The United Graduate School of Agricultural Sciences, Ehime University Satellite Business Venture, Ehime University Faculty of Agriculture, Ehime University

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  • In Vitro Protein Import of a Putative Amino Acid Transporter from Arabidopsis thaliana into Chloroplasts and Its Suborganellar Localization

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We identified a gene product of At5g19500 (At5g19500p) from Arabidopsis thaliana that is homologous to EcTyrP, a tyrosine-specific transporter from Escherichia coli. Computational analyses of the amino acid sequence of At5g19500p predicted 11 transmembrane domains (TMDs) and a potential plastid targeting signal at its amino terminus. As a first step toward understanding the possible role of At5g19500p in plant cells, we attempted to determine the localization of At5g19500p by an in vitro chloroplastic import assay using At5g19500p translated in a cell-free wheat germ system (Madin et al., Proc. Natl. Acad. Sci. USA, 97, 559–564 (2000)), followed by subfractionation of the chloroplasts. At5g19500p was successfully imported into chloroplasts, and the newly transported mature form of At5g19500p was recovered from the inner envelope membrane.

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