Lipid peroxidation is one of the consequences of environmental stress in plants and leads to the accumulation of highly toxic, reactive aldehydes. One of the processes to detoxify these aldehydes is their oxidation into carboxylic acids catalyzed by NAD(P)+-dependent ALDHs (aldehyde dehydrogenases). We investigated kinetic parameters of two Arabidopsis thaliana family 3 ALDHs, the cytosolic ALDH3H1 and the chloroplastic isoform ALDH3I1. Both enzymes had similar substrate specificity and oxidized saturated aliphatic aldehydes. Catalytic efficiencies improved with the increase of carbon chain length. Both enzymes were also able to oxidize α,β-unsaturated aldehydes, but not aromatic aldehydes. Activity of ALDH3H1 was NAD+-dependent, whereas ALDH3I1 was able to use NAD+ and NADP+. An unusual isoleucine residue within the coenzyme-binding cleft was responsible for the NAD+-dependence of ALDH3H1. Engineering the coenzyme-binding environment of ALDH3I1 elucidated the influence of the surrounding amino acids. Enzyme activities of both ALDHs were redox-sensitive. Inhibition was correlated with oxidation of both catalytic and non-catalytic cysteine residues in addition to homodimer formation. Dimerization and inactivation could be reversed by reducing agents. Mutant analysis showed that cysteine residues mediating homodimerization are located in the N-terminal region. Modelling of the protein structures revealed that the redox-sensitive cysteine residues are located at the surfaces of the subunits.
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Research Article|
February 24 2011
Engineering the nucleotide coenzyme specificity and sulfhydryl redox sensitivity of two stress-responsive aldehyde dehydrogenase isoenzymes of Arabidopsis thaliana
Naim Stiti;
Naim Stiti
*Institute of Molecular Physiology and Biotechnology of Plants, University of Bonn, Kirschallee 1, 53115 Bonn, Germany
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Isaac O. Adewale;
Isaac O. Adewale
†Department of Biochemistry, Obafemi Awolowo University, Ile-Ife, Osun, Nigeria
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Jan Petersen;
Jan Petersen
*Institute of Molecular Physiology and Biotechnology of Plants, University of Bonn, Kirschallee 1, 53115 Bonn, Germany
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Dorothea Bartels;
Dorothea Bartels
1
*Institute of Molecular Physiology and Biotechnology of Plants, University of Bonn, Kirschallee 1, 53115 Bonn, Germany
1Correspondence may be addressed to either of these authors (e-mail hhkirch@uni-bonn.de or dbartels@uni-bonn.de), who contributed equally to the work.
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Hans-Hubert Kirch
Hans-Hubert Kirch
1
*Institute of Molecular Physiology and Biotechnology of Plants, University of Bonn, Kirschallee 1, 53115 Bonn, Germany
1Correspondence may be addressed to either of these authors (e-mail hhkirch@uni-bonn.de or dbartels@uni-bonn.de), who contributed equally to the work.
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Publisher: Portland Press Ltd
Received:
August 23 2010
Revision Received:
December 15 2010
Accepted:
December 20 2010
Accepted Manuscript online:
December 20 2010
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2011 Biochemical Society
2011
Biochem J (2011) 434 (3): 459–471.
Article history
Received:
August 23 2010
Revision Received:
December 15 2010
Accepted:
December 20 2010
Accepted Manuscript online:
December 20 2010
Citation
Naim Stiti, Isaac O. Adewale, Jan Petersen, Dorothea Bartels, Hans-Hubert Kirch; Engineering the nucleotide coenzyme specificity and sulfhydryl redox sensitivity of two stress-responsive aldehyde dehydrogenase isoenzymes of Arabidopsis thaliana. Biochem J 15 March 2011; 434 (3): 459–471. doi: https://doi.org/10.1042/BJ20101337
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