A novel metalloproteinase present in freshly isolated rat glomeruli
| Autor: | Quoc C. Le, Sudhir V. Shah, Shirley L. Cortez, Huong T. Nguyen, William H. Baricos |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
food.ingredient
Physiology Renal glomerulus Kidney Glomerulus Matrix metalloproteinase Cell Fractionation Kidney Gelatin chemistry.chemical_compound food medicine Animals Gelatinase Trypsin Sodium dodecyl sulfate Edetic Acid Glycoproteins Gel electrophoresis Metalloproteinase Metalloendopeptidases Tissue Inhibitor of Metalloproteinases Hydrogen-Ion Concentration Rats Enzyme Activation Molecular Weight Dithiothreitol Biochemistry chemistry Electrophoresis Polyacrylamide Gel Phenanthrolines medicine.drug |
| Zdroj: | American Journal of Physiology-Renal Physiology. 260:F555-F561 |
| ISSN: | 1522-1466 1931-857X |
| DOI: | 10.1152/ajprenal.1991.260.4.f555 |
| Popis: | We have utilized [3H] gelatin to document high activity of a metalloproteinase present in freshly isolated rat glomeruli. [3H] gelatin degradation by glomeruli was markedly inhibited by EDTA (10 mM: -89 +/- 2.3%) and o-phenanthroline (2 mM: -72 +/- 0.1%), inhibitors of metalloproteinases. No significant inhibition of [3H]gelatin degradation was observed with inhibitors of serine or cysteine proteinases. Most (greater than 80%) of the glomerular metalloproteinase (GLOMP) activity was associated with the pellet after centrifugation of sonicated glomeruli at 100,000 g for 90 min. The pH optimum for gelatin degradation by sonicated glomeruli was approximately pH 8.5. Sodium dodecyl sulfate substrate (gelatin)-polyacrylamide gel electrophoresis revealed a single major band of EDTA-inhibitable gelatin-degrading activity with a molecular mass of approximately 116-125 kDa. The GLOMP activity was not inhibited by tissue inhibitors of metalloproteinases, did not appear to be latent, and was not activated by organomercurial activators of several latent metalloproteinases. GLOMP activity was increased 3.4-fold after incubation with trypsin (20 micrograms/ml, 25 min, 22 degrees C). These data indicate that GLOMP is distinct from the previously described matrix metalloproteinases, as well as other metalloproteinases present in the kidney, including the gelatinase secreted by cultured mesangial cells, Meprin, and endopeptidase 24.11 (enkephalinase, EC 3.4.24.11). |
| Databáze: | OpenAIRE |
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