Journal article
Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose
The plant glucan phosphatases Starch EXcess 4 (SEX4) and Like Sex Four2 (LSF2) apply different starch binding mechanisms. SEX4 contains a carbohydrate binding module, and LSF2 has two surface binding sites (SBSs). We determined KDapp for amylopectin and amylose, and KD for β-cyclodextrin and validated binding site mutants deploying affinity gel electrophoresis (AGE) and surface plasmon resonance.
SEX4 has a higher affinity for amylopectin; LSF2 prefers amylose and β-cyclodextrin. SEX4 has 50-fold lower KDapp for amylopectin compared to LSF2. Molecular dynamics simulations and AGE data both support long-distance mutual effects of binding at SBSs and the active site in LSF2.
| Language: | English |
|---|---|
| Publisher: | Wiley |
| Year: | 2016 |
| Pages: | 118-128 |
| ISSN: | 18733468 and 00145793 |
| Types: | Journal article |
| DOI: | 10.1002/1873-3468.12027 |
| ORCIDs: | Wilkens, Casper , Abou Hachem, Maher and Svensson, Birte |
Affinity gel electrophoresis Carbohydrate binding domain Like Sex Four2 Starch Excess 4 Surface binding sites Surface plasmon resonance
Amino Acid Substitution Amylopectin Amylose Arabidopsis Arabidopsis Proteins Binding Sites Carbohydrate Conformation Cytoplasmic Granules Dual-Specificity Phosphatases Kinetics LSF2 protein, Arabidopsis Models, Molecular Molecular Dynamics Simulation Mutation Plant Leaves Protein Conformation Recombinant Proteins SEX4 protein, Arabidopsis Substrate Specificity Surface Plasmon Resonance affinity gel electrophoresis beta-Cyclodextrins betadex carbohydrate binding domain surface binding sites surface plasmon resonance
