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Abstract
The accumulation of oxidatively damaged proteins is a well-known hallmark of aging and several neurodegenerative diseases including Alzheimer's, Parkinson's and Huntigton's diseases. These highly oxidized protein aggregates are in general not degradable by the main intracellular proteolytic machinery, the proteasomal system. One possible strategy to reduce the accumulation of such oxidized protein aggregates is the prevention of the formation of oxidized protein derivatives or to reduce the protein oxidation to a degree that can be handled by the proteasome. To do so an antioxidative strategy might be successful. Therefore, we undertook the present study to test whether antioxidants are able to prevent the protein oxidation and to influence the proteasomal degradation of moderate oxidized proteins. As a model protein we choose ferritin. H2O2 induced a concentration dependent increase of protein oxidation accompanied by an increased proteolytic susceptibility. This increase of proteolytic susceptibility is limited to moderate hydrogen peroxide concentrations, whereas higher concentrations are accompanied by protein aggregate formation.
Protective effects of the vitamin E derivative Trolox, the pyridoindole derivative Stobadine and of the standardized extracts of flavonoids from bark of Pinus Pinaster Pycnogenol® and from leaves of Ginkgo biloba (EGb 761) were studied on moderate damaged ferritin.
| Abbreviations | ||
| AAS | = | aminoadipic semialdehyde |
| AOx | = | antioxidant |
| BHT | = | butylated hydroxytoluole |
| BSA | = | bovine serum albumin |
| DEAE | = | diethylaminoethyl |
| DNPH | = | dinitrophenylhydrazine |
| EDTA | = | ethylenediaminetetraacetic acid |
| EGb 761 | = | Ginkgo biloba extract |
| ELISA | = | enzyme linked immunosorbent assey |
| FINH2 | = | fluoresceinamine |
| FPLC | = | fast protein liquid chromatography |
| Gb | = | Ginkgo biloba |
| GGS | = | γ-glutamic semialdehyde |
| MCO | = | metal catalyzed oxidation |
| PBS | = | phosphateous buffer saline |
| Pyc | = | Pycnogenol® |
| RNS | = | reactive nitrogen species |
| ROS | = | reactive oxygen species |
| SDS/PAGE | = | sodium-dodecyl-sulfate polyacrylamide gel electrophoresis |
| Sto | = | Stobadine |
| TCA | = | trichloroacetic acid |
| Tro | = | Trolox |
| Abbreviations | ||
| AAS | = | aminoadipic semialdehyde |
| AOx | = | antioxidant |
| BHT | = | butylated hydroxytoluole |
| BSA | = | bovine serum albumin |
| DEAE | = | diethylaminoethyl |
| DNPH | = | dinitrophenylhydrazine |
| EDTA | = | ethylenediaminetetraacetic acid |
| EGb 761 | = | Ginkgo biloba extract |
| ELISA | = | enzyme linked immunosorbent assey |
| FINH2 | = | fluoresceinamine |
| FPLC | = | fast protein liquid chromatography |
| Gb | = | Ginkgo biloba |
| GGS | = | γ-glutamic semialdehyde |
| MCO | = | metal catalyzed oxidation |
| PBS | = | phosphateous buffer saline |
| Pyc | = | Pycnogenol® |
| RNS | = | reactive nitrogen species |
| ROS | = | reactive oxygen species |
| SDS/PAGE | = | sodium-dodecyl-sulfate polyacrylamide gel electrophoresis |
| Sto | = | Stobadine |
| TCA | = | trichloroacetic acid |
| Tro | = | Trolox |