Purification and Molecular Properties of the Thylakoid-Bound Ascorbate Peroxidase in Spinach Chloroplasts
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概要
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The hydrogen peroxide that is photoproduced in thylakoids is scavenged by the thylakoid-bound ascorbate peroxidase (tAPX) [Miyake and Asada (1992) Plant Cell Physiol. 33: 541]. tAPX was purified from spinach thylakoids to homogeneity as judged by SDS-polyacrylamide gel electrophoresis, and its molecular properties were studied. Spinach tAPX was a monomer with a molecular weight of 40,000, which is about 10.000 higher than that of the stromal ascorbate peroxidase (sAPX) from spinach chloroplasts. tAPX cross-reacted with the antibody raised against sAPX from tea leaves, as determined by Western blotting, which also provided evidence for the higher molecular weight of tAPX from spinach thylakoids than that of tea sAPX. The amino acid sequence of the amino-terminal region of tAPX showed a low degree of homology to those of cylosolic APXs from spinach, pea and Arabidopsis thaliana, but a high degree of homology to that of stromal APX from tea. Thus, the amino-terminal region of tAPX seems not to be a domain required for binding of the enzyme to the thylakoid membranes. tAPX contained protoheme IX, as identified by its pyridine hemochromogen, and gave a Soret peak at 403 nm and 433 nm with an a band at 555 nm in its oxidized and reduced forms, respectively. Resembling sAPX but differing from cytosolic APX, tAPX showed high specificity for ascorbate as the electron donor. tAPX was inhibited by cyanide, thiol-modifying reagents, thiols and several suicide inhibitors, such as hydroxyurea and p-aminophenol.
- 日本植物生理学会の論文
著者
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Asada Kozi
The Research Institute for Food Science, Kyoto University
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Miyake Chikahiro
The Research Institute for Food Science, Kyoto University
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Cao W‐h
Inst. Genetics And Developmental Biology Chinese Acad. Sci. Beijing Chn
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Cao Wan-hong
The Research Institute For Food Science Kyoto University:(present)beijing Vegetable Research Center
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Miyake Chikahiro
The Research Institute For Food Science Kyoto University
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Asada Kozi
The Research Institute For Food Science Kyoto Niversity
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Asada Kozi
The Research Institulefor Fbod Science Kyoto University
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