Journal Article

In vivo Fragmentation of the Large Subunit of Ribulose-1,5-Bisphosphate Carboxylase by Reactive Oxygen Species in an Intact Leaf of Cucumber under Chilling-light Conditions

Ryouhei Nakano, Hiroyuki Ishida, Amane Makino and Tadahiko Mae

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 47, issue 2, pages 270-276
Published in print February 2006 | ISSN: 0032-0781
Published online February 2006 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci245
In vivo Fragmentation of the Large Subunit of Ribulose-1,5-Bisphosphate Carboxylase by Reactive Oxygen Species in an Intact Leaf of Cucumber under Chilling-light Conditions

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Previous studies have demonstrated that the large subunit (LSU) of ribulose-1,5-bisphosphate carboxylase (Rubisco) is site-specifically cleaved by a hydroxyl radical (·OH) generated in the illuminated chloroplast lysates or by an artificial ·OH-generating system. However, it is not known whether such cleavage of the LSU by reactive oxygen species (ROS) actually occurs in an intact leaf. When leaf discs of chilling-sensitive cucumber (Cucumis sativus L.) were illuminated at 4°C, five major fragments of the LSU were observed. This fragmentation was completely inhibited by ROS scavengers, such as n-propyl gallate (for ·OH) and 1,2-dihydroxybenzene-3,5-disulfonic acid (Tiron) (for superoxide). FeSO4 stimulated this fragmentation, whereas an iron-specific chelator, deferoxamine, suppressed it. Furthermore, such fragments were identical to those generated from the purified Rubisco by an ·OH-generating system in vitro on two-dimensional PAGE. These results indicate that the direct fragmentation of the LSU by reacive oxygen species also occurs in an intact leaf.

Keywords: Chilling; Cucumber (Cucumis sativus L.); Photoinhibiton; PSI; Reactive oxygen species; Rubisco; APX, ascorbate peroxidase; DTT, dithiothreitol; ·OH, hydroxyl radical; LSU, large subunit of Rubisco; ROS, reactive oxygen species; Rubisco, ribulose-1,5-bisphosphate carboxylase/oxygensase; SOD, superoxide dismutase; Tiron, 1,2-dihydroxybenzene-3,5-disulfonic acid

Journal Article.  5505 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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