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 validat...

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Bibliographic Details
Published in:FEBS letters 2016-01, Vol.590 (1), p.118-128
Main Authors: Wilkens, Casper, Auger, Kyle D., Anderson, Nolan T., Meekins, David A., Raththagala, Madushi, Abou Hachem, Maher, Payne, Christina M., Gentry, Matthew S., Svensson, Birte, Fluegge, Ulf‐Ingo
Format: Article
Language:English
Subjects:
affinity gel electrophoresis
Amino Acid Substitution
Amylopectin - chemistry
Amylopectin - metabolism
Amylose - chemistry
Amylose - metabolism
Arabidopsis - enzymology
Arabidopsis Proteins - chemistry
Arabidopsis Proteins - genetics
Arabidopsis Proteins - metabolism
beta-Cyclodextrins - chemistry
beta-Cyclodextrins - metabolism
Binding Sites
carbohydrate binding domain
Carbohydrate Conformation
Cytoplasmic Granules - chemistry
Cytoplasmic Granules - enzymology
Cytoplasmic Granules - metabolism
Dual-Specificity Phosphatases - chemistry
Dual-Specificity Phosphatases - genetics
Dual-Specificity Phosphatases - metabolism
Kinetics
Like Sex Four2
Models, Molecular
Molecular Dynamics Simulation
Mutation
Plant Leaves - enzymology
Protein Conformation
Recombinant Proteins - chemistry
Recombinant Proteins - metabolism
Starch Excess 4
Substrate Specificity
surface binding sites
Surface Plasmon Resonance
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Summary: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.
ISSN:0014-5793
1873-3468
DOI:10.1002/1873-3468.12027