Comprehensive survey of proteins targeted by chloroplast thioredoxin
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- Ken Motohashi
- Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan; and Core Research for Evolutional Science and Technology (CREST), Genetic Programming Team 13, Nogawa 907, Miyamae-ku, Kawasaki 216-0001, Japan
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- Aiko Kondoh
- Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan; and Core Research for Evolutional Science and Technology (CREST), Genetic Programming Team 13, Nogawa 907, Miyamae-ku, Kawasaki 216-0001, Japan
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- Michael T. Stumpp
- Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan; and Core Research for Evolutional Science and Technology (CREST), Genetic Programming Team 13, Nogawa 907, Miyamae-ku, Kawasaki 216-0001, Japan
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- Toru Hisabori
- Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan; and Core Research for Evolutional Science and Technology (CREST), Genetic Programming Team 13, Nogawa 907, Miyamae-ku, Kawasaki 216-0001, Japan
説明
<jats:p> Possible target proteins of chloroplast thioredoxin (Trx) have been investigated in the stroma lysate of spinach chloroplasts. For that purpose, we immobilized a mutant of <jats:italic>m</jats:italic> -type Trx in which an internal cysteine at the active site was substituted with serine, on cyanogen bromide-activated resin. By using this resin, the target proteins in chloroplast were efficiently acquired when they formed the mixed-disulfide intermediates with the immobilized Trxs. We could acquire Rubisco activase (45 kDa) and 2-Cys-type peroxiredoxin (Prx), which were recently identified as targets of chloroplast Trxs. Glyceraldehyde-3-phosphate dehydrogenase and sedoheputulose 1,7- <jats:italic>bis</jats:italic> phosphatase, well-known thiol enzymes in the Calvin cycle, also were recognized among the collected proteins, suggesting the method is applicable for our purpose. Furthermore, four proteins were identified from a homology search of the NH <jats:sub>2</jats:sub> -terminal sequence of the acquired proteins: glutamine synthetase, a protein homologous to chloroplast cyclophilin, a homolog of Prx-Q, and the Rubisco small subunit. The Trx susceptibilities of the recombinant cyclophilin and Prx-Q of <jats:italic>Arabidopsis thaliana</jats:italic> were then examined. The method developed in the present study is thus applicable to investigate the various redox networks via Trxs and the related enzymes in the cell. </jats:p>
収録刊行物
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- Proceedings of the National Academy of Sciences
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Proceedings of the National Academy of Sciences 98 (20), 11224-11229, 2001-09-11
Proceedings of the National Academy of Sciences
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キーワード
- Models, Molecular
- Chloroplasts
- Ribulose-Bisphosphate Carboxylase
- Molecular Sequence Data
- Arabidopsis
- Protein Structure, Secondary
- Cyclophilins
- Thioredoxins
- Glutamate-Ammonia Ligase
- Spinacia oleracea
- Serine
- Amino Acid Sequence
- Cysteine
- Disulfides
- Cloning, Molecular
- Plant Proteins
- Binding Sites
- Glyceraldehyde-3-Phosphate Dehydrogenases
- Peroxiredoxins
- Biological Sciences
- Phosphoric Monoester Hydrolases
- Recombinant Proteins
- Kinetics
- Protein Subunits
- Amino Acid Substitution
- Peroxidases
- Mutagenesis, Site-Directed
- Resins, Plant
詳細情報 詳細情報について
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- CRID
- 1361137044762566400
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- NII論文ID
- 80015134224
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- ISSN
- 10916490
- 00278424
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- PubMed
- 11553771
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- データソース種別
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- Crossref
- CiNii Articles
- OpenAIRE