A soybean sucrose binding protein independently mediates nonsaturable sucrose uptake in yeast.
Heterologous expression of a cDNA encoding a 62-kD soybean sucrose binding protein in yeast demonstrates that this protein, independent of other plant proteins, mediates sucrose uptake across the plasma membrane. Sucrose binding protein-mediated sucrose uptake is nonsaturable up to 30 mM sucrose, is specific for sucrose, and is relatively insensitive to treatment with sulfhydryl-modifying reagents. Alteration of the external pH or pretreatment of the yeast cells with protonophores did not significantly affect the rate of 14C-sucrose uptake. This demonstrates that sucrose binding protein-mediated sucrose uptake is not dependent on H+ movement and delineates it from other plant sucrose transporters. Physiological characterization of sucrose uptake into higher plant cells has shown the presence of both saturable and nonsaturable uptake components. The nonsaturable mechanism is relatively insensitive to external pH, pretreatment with protonophores, and treatment with sulfhydryl-modifying reagents. Sucrose binding protein-mediated sucrose uptake in yeast mimics this physiologically described, but mechanistically undefined, nonsaturable sucrose uptake mechanism in higher plants. Functional characterization of the sucrose binding protein thus defines both a novel component of sucrose uptake and provides important insight into this nonsaturable sucrose uptake mechanism, which has remained enigmatic since its physiological description.
Overvoorde
P J
PJ
Department of Botany, Washington State University, Pullman 99164-4238, USA.
Frommer
W B
WB
Grimes
H D
HD
D019297:2,4-Dinitrophenol; D001483:Base Sequence; D001692:Biological Transport; D002250:Carbon Radioisotopes; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002352:Carrier Proteins / Q000096:biosynthesis / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004140:Dinitrophenols / Q000494:pharmacology; D005838:Genotype; D007700:Kinetics; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D037121:Plant Lectins; D010940:Plant Proteins / Q000096:biosynthesis / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D030262:Soybean Proteins; D013025:Glycine max / Q000378:metabolism*; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D018076:DNA, Complementary; D004140:Dinitrophenols; D026901:Membrane Transport Proteins; D009838:Oligodeoxyribonucleotides; D037121:Plant Lectins; D010940:Plant Proteins; D011994:Recombinant Proteins; D030262:Soybean Proteins; D013439:Sulfhydryl Reagents; C078849:SBP protein, Glycine max; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D013395:Sucrose; D019297:2,4-Dinitrophenol