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J. Biol. Chem., Vol. 277, Issue 21, 18373-18382, May 24, 2002
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From the Betaine is an important osmoprotectant in many
plants, but its transport activity has only been demonstrated using a
proline transporter from tomato, a betaine-nonaccumulating 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/4-aminobutyrate transporters from mammalian cells
were low but were high to proline transporters from
Arabidopsis and tomato. Two full-length transporters could
complement the Na+-sensitive phenotype of the
Escherichia 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 of protein). The
uptakes of betaine and proline were significantly inhibited by mono-
and dimethylglycine but only partially inhibited by betaine aldehyde, choline, and 4-aminobutyrate. 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 Thr290-Thr-Ser292 in a
putative periplasmic loop to Arg290-Gly-Arg292
yielded the active transporter independent of salts, suggesting the
positive charge induced a conformational change to the 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.
The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBankTM/EBI Data Bank with accession number(s) AB075902, AB075903, and AB075904.
Functional Characterization of Betaine/Proline Transporters in
Betaine-accumulating Mangrove*
§,
,,¶¶
Research Institute, ¶ Faculty of
Science and Technology, and ** School of Agriculture, Meijo
University, Nagoya 468-8502, Japan, § Department of
Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok
10330, Thailand, Faculty of Pharmaceutical Science, Chiba
University, Chiba 263-8522, Japan, Faculty
of Agriculture, University of the Ryukyus, Okinawa 903-0213, Japan, and
§§ Graduate School of Agricultural Science,
Nagoya University, Chikusa-ku, Nagoya, Aichi 464-8601 Japan
*
This work was supported in part by grants-in-aid for
scientific research from the Ministry of Education and Science and
Culture of Japan and from the High-Tech Research Center of Meijo
University.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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