Characterization and purification of a lipoxygenase inhibitor in human epidermoid carcinoma A431 cells
| Autor: | Chia-Yi Yang, Yu-Tsong Lee, Hui-Sheng Huang, Wen Chang Chang, Ching-Jiunn Chen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1997 |
| Předmět: |
Lipoxygenase
Iodoacetates Biochemistry chemistry.chemical_compound Cytosol Microsomes Enzyme Stability medicine Tumor Cells Cultured Humans Isoelectric Point Lipoxygenase Inhibitors Molecular Biology chemistry.chemical_classification Arachidonic Acid biology Glutathione peroxidase Temperature Cell Biology Glutathione Trypsin Chromatography Ion Exchange Iodoacetic Acid Molecular Weight chemistry Epidermoid carcinoma Sephadex biology.protein Chromatography Gel Electrophoresis Polyacrylamide Gel A431 cells medicine.drug Research Article |
| Popis: | A lipoxygenase inhibitor in the cytosolic fraction of human epidermoid carcinoma A431 cells was characterized and purified. The cytosolic inhibitor lost the inhibitory activity upon heating at 75 degrees C for 15 min or pretreating with 1 mg/ml trypsin at 37 degrees C for 60 min. Cytosol, after dialysis, lost the inhibitory activity but its inhibitory activity recovered when 1 mM GSH was added to the dialysate. The inhibitory activity of cytosol was also abolished by treatment either with 1 mM iodoacetate at 4 degrees C for 1 h or with 0.5 mM H2O2. The pI of the inhibitor was approx. 7.0. In addition to 12-lipoxygenase, the inhibitor inhibited the activities of 5-lipoxygenase and fatty acid cyclo-oxygenase in a cell-free system. The inhibitor was purified by a series of column chromatographies using CM Sephadex C-50, Sephadex G-100 SF and Mono P columns. A major 22 kDa protein was obtained that was distinct from selenium-dependent glutathione peroxidase. |
| Databáze: | OpenAIRE |
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