Oxidation of hydroquinone by both cellular and extracellular grapevine peroxidase fractions.
The oxidation of hydroquinone by two peroxidase (EC 1.11.1.7) fractions obtained from the cells and spent medium of cell cultures of grapevine (Vitis vinifera cv Monastrell) has been studied, and their comparative efficacy (kcat/KM ratio) studied in both the H2O2-consuming and hydroquinone-consuming reactions. While the efficacy in the H2O2-consuming reaction is practically identical for both enzyme fractions, the cellular peroxidase has five-fold more efficacy in the hydroquinone-consuming reaction than the peroxidase located in the spent medium. Screening of cellular peroxidases capable of oxidizing hydroquinone on polyacrylamide gels, by means of a staining reaction based on the nucleophilic attack of 4-aminoantipyrine on p-benzoquinone in acidic media, reveals that all the cellular peroxidase isoenzymes are capable of oxidizing hydroquinone, probably yielding a quinone-diimine as a product of the staining reaction. Since isoperoxidases found in cellular fractions are also present in the spent medium, the values found for the different efficacies in the hydroquinone-consuming reaction must be considered as the results of the different proportions in which each peroxidase isoenzyme was found in the two fractions. The localization of a benzoquinone-generating system of high efficacy inside the plant cell, and probably located in vacuoles, is discussed with respect to the harmful role which the quinone/semiquinone pair might play in cell death, as part of the hypersensitive response expressed within the mechanism of plant disease resistance.
Zapata
J M
JM
Department of Plant Biology (Plant Physiology), University of Murcia, Spain.
Caldéron
A A
AA
Muñoz
R
R
Ros Barceló
A
A
D000675:Ampyrone / Q000378:metabolism; D016227:Benzoquinones / Q000378:metabolism; D002478:Cells, Cultured; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006873:Hydroquinones / Q000378:metabolism*; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D010944:Plants / Q000201:enzymology*
D016227:Benzoquinones; D006873:Hydroquinones; D007527:Isoenzymes; D000675:Ampyrone; C004532:quinone; D006861:Hydrogen Peroxide; D009195:Peroxidase; C031927:hydroquinone