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M112012200v1
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Papers In Press, published online ahead of print March 20, 2002
J. Biol. Chem, 10.1074/jbc.M112012200
Submitted on December 17, 2001
Revised on March 20, 2002
Accepted on March 20, 2002

Functional characterization of Betaine/proline transporters in betaine accumulating mangrove

Rungaroon Waditee, Takashi Hibino, Yoshito Tanaka, Tatsunosuke Nakamura, Aran Incharoensakdi, Shinsuke Hayakawa, Shigetoshi Suzuki, Yuzo Futsuhara, Yoshinobu Kawamitsu, Tetsuko Takabe, and Teruhiro Takabe

Research Institute, Meijo University, Nagoya, Aichi 468-8502

Corresponding Author: takabe{at}ccmfs.meijo-u.ac.jp

Betaine is an important osmoprotectant in many plants, but its transport activity has only been demonstrated using a proline transporter from tomato, a betaine non accumulating plant. In this study, two full length and one partial transporter genes were isolated from betaine accumulating mangrove Avicennia marina. Their homologies to betaine transporters from bacteria and betaine/GABA transporters from mammalian cells were low, but high to proline transporters from Arabidopsis and tomato. Two full length transporters could complement the Na+-sensitive phenotype of the E. coli mutant deficient in betT, putPA, proP, and proU. Both transporters could efficiently take up betaine and proline with similar affinities (Km, 0.32-0.43 mM) and maximum velocities (1.9-3.6 nmol/min mg protein). The uptakes of betaine and proline were significantly inhibited by mono- and di-methylglycine, but only partially inhibited by betaine aldehyde, choline, and GABA. Sodium- and potassium-chloride markedly enhanced betaine uptake rates with optimum concentrations at 0.5 M whereas sucrose showed only modest activation. The change of amino acids 290Thr-Thr-Ser292 in a putative periplasmic loop to 290Arg-Gly-Arg292 yielded the active transporter independent of salts, suggesting the positive charge induced conformational change to active form. These data clearly indicate that the betaine accumulating mangrove contains betaine/proline transporters whose properties are distinct from betaine transporters of bacteria and mammalian cells.


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