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

University of Kentucky

Research output: Contribution to journal › Journal article › Research › peer-review

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 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 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
Journal	FEBS Letters
Volume	590
Issue number	1
Pages (from-to)	118-128
Number of pages	11
ISSN	0014-5793
DOIs	https://doi.org/10.1002/1873-3468.12027
Publication status	Published - 2016

Keywords

Affinity gel electrophoresis
Carbohydrate binding domain
Like Sex Four2
Starch Excess 4
Surface binding sites
Surface plasmon resonance

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10.1002/1873-3468.12027

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@article{c9941911c21d44988509a8444fd56a22,

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 = "Affinity gel electrophoresis, Carbohydrate binding domain, Like Sex Four2, Starch Excess 4, 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",

year = "2016",

doi = "10.1002/1873-3468.12027",

language = "English",

volume = "590",

pages = "118--128",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "John Wiley and Sons Inc.",

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TY - JOUR

T1 - Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

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

Y1 - 2016

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.

AB - 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.

KW - Affinity gel electrophoresis

KW - Carbohydrate binding domain

KW - Like Sex Four2

KW - Starch Excess 4

KW - Surface binding sites

KW - Surface plasmon resonance

U2 - 10.1002/1873-3468.12027

DO - 10.1002/1873-3468.12027

M3 - Journal article

C2 - 26763114

SN - 0014-5793

VL - 590

SP - 118

EP - 128

JO - FEBS Letters

JF - FEBS Letters

IS - 1

ER -

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

Abstract

Keywords

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