Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

Casper Wilkens; Kyle D. Auger; Nolan T. Anderson; David A. Meekins; Madushi Raththagala; Maher Abou Hachem; Christina M. Payne; Matthew S. Gentry; Birte Svensson

doi:10.1002/1873-3468.12027

Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

Casper Wilkens, Kyle D. Auger, Nolan T. Anderson, David A. Meekins, Madushi Raththagala, Maher Abou Hachem, Christina M. Payne, Matthew S. Gentry, Birte Svensson

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

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 K_Dapp for amylopectin and amylose, and K_D 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 K_Dapp 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.

Original language	English
Pages (from-to)	118-128
Number of pages	11
Journal	FEBS Letters
Volume	590
Issue number	1
DOIs	https://doi.org/10.1002/1873-3468.12027
State	Published - Jan 2016

Bibliographical note

Publisher Copyright:
© 2015 Federation of European Biochemical Societies.

Funding

This work was supported by The Danish Council for Independent Research | Natural Sciences (FNU) to the project ‘Discovery and Characterization of Carbohydrate Surface Binding Sites (SBS) in Polysaccharide Converting Enzymes’ and the Biacore T100 instrument (both to BS) and a National Science Foundation CAREER Grant MCB-1252345, National Institutes of Health Grant P20GM103486, Kentucky Science and Energy Foundation Grant KSE-2971-RDE-0147, Mizutani Foundation for Glycoscience Award, and NSF Grant IIA-1355438 (all to MSG). The Technical University of Denmark co-financed a PhD fellowship (CW). Computational resources were provided by the University of Kentucky and the Extreme Science and Engineering Discovery Environment (XSEDE) supported by National Science Foundation grant number ACI-1053575 (allocation TG-MCB090159). Andreas Blennow, University of Copenhagen is thanked for fruitful discussions on the structure of phosphorylated starch and David Wilson, Cornell University for the T. fusca Cel6A.

Funders	Funder number
National Science Foundation (NSF)	IIA-1355438, MCB-1252345
National Institutes of Health (NIH)
National Institute of General Medical Sciences	P20GM103486
Kentucky Science and Energy Foundation	KSE-2971-RDE-0147
Cornell High Energy Synchrotron Source, Cornell University
University of Kentucky	TG-MCB090159, ACI-1053575
Natur og Univers, Det Frie Forskningsråd
Mizutani Foundation for Glycoscience
Danmarks Tekniske Universitet

Keywords

Like Sex Four2
Starch Excess 4
affinity gel electrophoresis
carbohydrate binding domain
surface binding sites
surface plasmon resonance

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1002/1873-3468.12027

Cite this

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title = "Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose",

abstract = "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.",

keywords = "Like Sex Four2, Starch Excess 4, affinity gel electrophoresis, carbohydrate binding domain, surface binding sites, surface plasmon resonance",

author = "Casper Wilkens and Auger, {Kyle D.} and Anderson, {Nolan T.} and Meekins, {David A.} and Madushi Raththagala and {Abou Hachem}, Maher and Payne, {Christina M.} and Gentry, {Matthew S.} and Birte Svensson",

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year = "2016",

month = jan,

doi = "10.1002/1873-3468.12027",

language = "English",

volume = "590",

pages = "118--128",

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AU - Wilkens, Casper

AU - Auger, Kyle D.

AU - Anderson, Nolan T.

AU - Meekins, David A.

AU - Raththagala, Madushi

AU - Abou Hachem, Maher

AU - Payne, Christina M.

AU - Gentry, Matthew S.

AU - Svensson, Birte

PY - 2016/1

Y1 - 2016/1

N2 - 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.

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KW - Like Sex Four2

KW - Starch Excess 4

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KW - carbohydrate binding domain

KW - surface binding sites

KW - surface plasmon resonance

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JO - FEBS Letters

JF - FEBS Letters

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Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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