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Reference Detail
| Reference | ||
|---|---|---|
| Author | Reinders A., Sun Y., Karvonen K.L., Ward J.M. | |
| Title | Identification of amino acids important for substrate specificity in sucrose transporters using gene shuffling. | |
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Abstract: Plant sucrose transporters (SUTs) are H+-coupled uptake transporters. Type I and II (SUTs) are phylogenetically related but have different substrate specificities. Type I SUTs transport sucrose, maltose and a wide range of natural and synthetic alpha- and beta-glucosides. Type II SUTs are more selective for sucrose and maltose. Here, we investigated the structural basis for this difference in substrate specificity. We used a novel gene shuffling method called Synthetic Template Shuffling (STS) to introduce 62 differentially conserved amino acid residues from type I SUTs into OsSUT1, a type II SUT from rice. The OsSUT1 variants were tested for their ability to transport the fluorescent coumarin beta-glucoside esculin when expressed in yeast. Fluorescent yeast cells were selected using fluorescence-activated cell sorting (FACS). Substitution of five amino acids present in type I SUTs in OsSUT1 was found to be sufficient to confer esculin uptake activity. The changes clustered in two areas of the OsSUT1 protein, in the first loop and the top of TMS2 (T80L, A86K) and in TMS5 (S220A, S221A, T224Y). The substrate specificity of this OsSUT1 variant was almost identical to that of type I SUTs. Corresponding changes in the sugarcane type II transporter ShSUT1 also changed substrate specificity indicating that these residues contribute to substrate specificity in type II SUTs in general. |
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| Journal | J. Biol. Chem. | |
| Country | USA | |
| Volume | 287(36) | |
| Pages | 30296-304 | |
| Year | 2012 | |
| PubMed ID | 22807445 | |
| PubMed Central ID | - | |
| DOI | - | |
| URL | - | |
| Relation | ||
| Gene | SUT1 | |
| INSD | - | |
| Strain | Wild Core Collection | - |
| Induced Mutation Lines(NIG Collection) | - | |
| Sterile Seed Strain | - | |
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Lethal Embryo Mutantion Strain |
- | |
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Stages in Each Organ - Muant Lines (Gene) |
- | |
| Cultivated Varieties(NIG Collection) | - | |
| Stages in Each Organ | - | |
