Assay of DNA Photolyase Activity in Spinach Leaves in Relation to Cell Compartmentation-Evidence for Lack of DNA Photolyase in Chloroplasts
| Autor: | Jorn Goss, Megumi Hada, Masateru Shin, Keisuke Hino, Gunther Buchholz, Eckard Wellmann |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Chloroplasts
DNA Repair DNA repair education Enzyme-Linked Immunosorbent Assay Pyrimidine dimer Applied Microbiology and Biotechnology Biochemistry Analytical Chemistry Deoxyribodipyrimidine photo-lyase Mice chemistry.chemical_compound Spinacia oleracea Animals Photolyase Molecular Biology biology Organic Chemistry Antibodies Monoclonal food and beverages General Medicine DNA photolyase biology.organism_classification Molecular biology Cell Compartmentation Plant Leaves chemistry biology.protein Spinach Deoxyribodipyrimidine Photo-Lyase DNA Biotechnology Peroxidase |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 64:1288-1291 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1271/bbb.64.1288 |
| Popis: | Spinach cyclobutane pyrimidine dimer (CPD)-specific DNA photolyase was successfully detected in leaf extracts by an assay system for plant photolyase using an improved enzyme-linked immunosorbent assay (ELISA) which was newly introduced by novel horseradish peroxidase (HRP)-linked CPD specific monoclonal antibodies. The assay system includes two main steps: a photorepair reaction of CPD introduced in substrate DNA and measurement of CPD remained after the photorepair by the improved ELISA. When CPD- induced salmon sperm DNA was used as a substrate, high CPD-photolyase activities were observed in the enzyme fraction prepared from whole spinach leaf extracts, but not from chloroplast extracts. This strongly suggests that spinach CPD-specific photolyases are localized in cell compartments other than chloroplasts. |
| Databáze: | OpenAIRE |
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