Identification of key residues for pH dependent activation of violaxanthin de-epoxidase from Arabidopsis thaliana.

PloS One
Christian FufezanTomas Morosinotto

Abstract

Plants are often exposed to saturating light conditions, which can lead to oxidative stress. The carotenoid zeaxanthin, synthesized from violaxanthin by Violaxanthin De-Epoxidase (VDE) plays a major role in the protection from excess illumination. VDE activation is triggered by a pH reduction in the thylakoids lumen occurring under saturating light. In this work the mechanism of the VDE activation was investigated on a molecular level using multi conformer continuum electrostatic calculations, site directed mutagenesis and molecular dynamics. The pK(a) values of residues of the inactive VDE were determined to identify target residues that could be implicated in the activation. Five such target residues were investigated closer by site directed mutagenesis, whereas variants in four residues (D98, D117, H168 and D206) caused a reduction in enzymatic activity indicating a role in the activation of VDE while D86 mutants did not show any alteration. The analysis of the VDE sequence showed that the four putative activation residues are all conserved in plants but not in diatoms, explaining why VDE in these algae is already activated at higher pH. Molecular dynamics showed that the VDE structure was coherent at pH 7 with a low amount ...Continue Reading

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Citations

Nov 2, 2013·Chemical Reviews·Alexander R MoiseEleanore T Wurtzel
Jan 27, 2016·Journal of Plant Physiology·Zhen-Hui Kang, Gui-Xue Wang
Jul 24, 2013·Biophysical Chemistry·Diana SimionatoTomas Morosinotto
Dec 3, 2014·Cell Calcium·Giovanni FinazziDaphné Seigneurin-Berny
Jul 31, 2013·Progress in Lipid Research·Ildikó DomonkosBettina Ughy
Mar 25, 2017·Journal of Experimental Botany·Ildikò Szabò, Cornelia Spetea
May 20, 2020·Frontiers in Plant Science·Reimund Goss, Dariusz Latowski
Feb 23, 2021·Frontiers in Genetics·Susana González-MoralesAdalberto Benavides-Mendoza

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Methods Mentioned

BETA
x-ray crystallography

Software Mentioned

Multi Conformer Continuum Electrostatic ( MCCE2
DELPHI
Python
MCCE
ClustalW
Gromacs
p3d
mfe
py

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