Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals

Publication: Research - peer-reviewJournal article – Annual report year: 2001

Standard

Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals. / Galuszka, P.; Frebort, I.; Sebela, M.; Sauer, P.; Jacobsen, Susanne; Pec, P.

In: European Journal of Biochemistry, Vol. 268, No. 2, 2001, p. 450-461.

Publication: Research - peer-reviewJournal article – Annual report year: 2001

Harvard

Galuszka, P, Frebort, I, Sebela, M, Sauer, P, Jacobsen, S & Pec, P 2001, 'Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals' European Journal of Biochemistry, vol 268, no. 2, pp. 450-461.

APA

Galuszka, P., Frebort, I., Sebela, M., Sauer, P., Jacobsen, S., & Pec, P. (2001). Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals. European Journal of Biochemistry, 268(2), 450-461.

CBE

MLA

Vancouver

Author

Galuszka, P.; Frebort, I.; Sebela, M.; Sauer, P.; Jacobsen, Susanne; Pec, P. / Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals.

In: European Journal of Biochemistry, Vol. 268, No. 2, 2001, p. 450-461.

Publication: Research - peer-reviewJournal article – Annual report year: 2001

Bibtex

@article{9c17cf8b0bec4158a41d8505691152b0,
title = "Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals",
keywords = "protein",
publisher = "Springer Verlag",
author = "P. Galuszka and I. Frebort and M. Sebela and P. Sauer and Susanne Jacobsen and P. Pec",
year = "2001",
volume = "268",
number = "2",
pages = "450--461",
journal = "European Journal of Biochemistry",
issn = "0014-2956",

}

RIS

TY - JOUR

T1 - Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals

A1 - Galuszka,P.

A1 - Frebort,I.

A1 - Sebela,M.

A1 - Sauer,P.

A1 - Jacobsen,Susanne

A1 - Pec,P.

AU - Galuszka,P.

AU - Frebort,I.

AU - Sebela,M.

AU - Sauer,P.

AU - Jacobsen,Susanne

AU - Pec,P.

PB - Springer Verlag

PY - 2001

Y1 - 2001

N2 - An enzyme degrading cytokinins with isoprenoid side chain, previously named cytokinin oxidase, was purified to near homogeneity from wheat and barley grains. New techniques were developed for the enzyme activity assay and staining on native electrophoretic gels to identify the protein. The purified wheat enzyme is a monomer 60 kDa, its N-terminal amino-acid sequence shows similarity to hypothetical cytokinin oxidase genes from Arabidopsis thaliana, but not to the enzyme from maize. N-6-isopentenyl-2-(2-hydroxyethylamino)-9-methyladenine is the best substrate from all the cytokinins tested. Interestingly, oxygen was not required and hydrogen peroxide not produced during the catalytic reaction, so the enzyme behaves as a dehydrogenase rather than an oxidase. This was confirmed by the ability of the enzyme to transfer electrons to artificial electron acceptors, such as phenazine methosulfate and 2,6-dichlorophenol-indophenol. 2,3-Dimethoxy-5-methyl-1,4- benzoquinone, a precursor of the naturally occurring electron acceptor ubiquinone, readily interacts with the enzyme in micromolar concentrations. Typical flavoenzyme inhibitors such as acriflavine and diphenyleneiodonium inhibited this enzyme activity. Presence of the flavin cofactor in the enzyme was confirmed by differential pulse polarography and by measuring the fluorescence emission spectrum. Possible existence of a second redox centre is discussed.

AB - An enzyme degrading cytokinins with isoprenoid side chain, previously named cytokinin oxidase, was purified to near homogeneity from wheat and barley grains. New techniques were developed for the enzyme activity assay and staining on native electrophoretic gels to identify the protein. The purified wheat enzyme is a monomer 60 kDa, its N-terminal amino-acid sequence shows similarity to hypothetical cytokinin oxidase genes from Arabidopsis thaliana, but not to the enzyme from maize. N-6-isopentenyl-2-(2-hydroxyethylamino)-9-methyladenine is the best substrate from all the cytokinins tested. Interestingly, oxygen was not required and hydrogen peroxide not produced during the catalytic reaction, so the enzyme behaves as a dehydrogenase rather than an oxidase. This was confirmed by the ability of the enzyme to transfer electrons to artificial electron acceptors, such as phenazine methosulfate and 2,6-dichlorophenol-indophenol. 2,3-Dimethoxy-5-methyl-1,4- benzoquinone, a precursor of the naturally occurring electron acceptor ubiquinone, readily interacts with the enzyme in micromolar concentrations. Typical flavoenzyme inhibitors such as acriflavine and diphenyleneiodonium inhibited this enzyme activity. Presence of the flavin cofactor in the enzyme was confirmed by differential pulse polarography and by measuring the fluorescence emission spectrum. Possible existence of a second redox centre is discussed.

KW - protein

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

SN - 0014-2956

IS - 2

VL - 268

SP - 450

EP - 461

ER -