Time course of release into the medium of newly synthesized proteins by cultured aortic endothelial cells. Role of serum in preventing proteolytic degradation
| Autor: | E S Dickinson, P A Dennis, L L Slakey |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
Time Factors
Immunoprecipitation Swine medicine.medical_treatment Period (gene) Peptidyl-Dipeptidase A chemistry.chemical_compound medicine Animals Protease Inhibitors Endothelium Aorta Cells Cultured chemistry.chemical_classification Protease biology Immune Sera Leupeptin Proteins Molecular biology Macroglobulin Fibronectin Enzyme Biochemistry chemistry biology.protein Cardiology and Cardiovascular Medicine Pepstatin |
| Zdroj: | Arteriosclerosis (Dallas, Tex.). 6(6) |
| ISSN: | 0276-5047 |
| Popis: | Endothelial cells from pig aortas were labeled with 35S-methionine, and the soluble proteins that were released into the culture medium were examined by SDS-PAGE. Proteins were collected during labeling, and during the intervals 0 to 3, 3 to 6, 6 to 12, and 12 to 24 hours of chase after a 1-hour labeling period, and during the intervals 0 to 12, 12 to 24, and 24 to 48 hours of chase after a 24-hour labeling period. Release of radiolabeled soluble protein from cells into the medium continued over the longest time periods examined. If the cells were labeled for 1 hour, characteristic patterns of proteins appeared in the medium during the labeling period, early (0- to 3-hour postlabeling) and late (6- to 24-hour postlabeling). The time course of release of fibronectin (Fn) and of angiotensin-converting enzyme (ACE) was determined by using immunoprecipitation. ACE appeared only after 6 hours of chase. Fn appeared throughout the chase period. The effects of serum and of the protease inhibitors alpha-2 macroglobulin, pepstatin, and leupeptin on protein release were examined. In the absence of serum, endothelial cell culture medium contained substantial protease activity capable of completely degrading most of the released proteins; a major effect of serum was to protect newly released protein from degradation. |
| Databáze: | OpenAIRE |
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