In vitro Ca deposition by rat matrix vesicles: is the membrane association of alkaline phosphatase essential for matrix vesicle-mediated calcium deposition?
| Autor: | Lori L. Davis, D.C. Morris, Paul E. Moylan, Clarke H. Anderson, Howard H.T. Hsu |
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| Rok vydání: | 1993 |
| Předmět: |
Gel electrophoresis
medicine.diagnostic_test Phosphoric Diester Hydrolases Vesicle Phosphatidylinositol Diacylglycerol-Lyase chemistry.chemical_element Calcium Matrix (biology) In Vitro Techniques Alkaline Phosphatase Biochemistry Extracellular Matrix Rats Membrane Phosphoinositide Phospholipase C chemistry Western blot Solubility Liposomes medicine Alkaline phosphatase Animals Specific activity |
| Zdroj: | The International journal of biochemistry. 25(12) |
| ISSN: | 0020-711X |
| Popis: | 1. 1. Phosphatidylinositol phospholipase C (PI-PLC) treatment of rachitic rat matrix vesicles (MVs) released about 80% of membrane-bound alkaline phosphatase (ALP), AMPase, PPiase into the media. 2. 2. About 20% hydrolytic activity was not released from MV membranes by PI-PLC treatment. 3. 3. SDS-polyacrylamide gel electrophoresis and Western blot analysis showed only one immunoreactive protein corresponding to the molecular weight of ALP present in the soluble fraction after PI-PLC treatment. 4. 4. The specific activity of the released ALP was at least 5-fold higher than the residual activity. 5. 5. After PI-PLC treatment, MVs also demonstrated an 80% reduction of AMP- or βGP-dependent calcium deposition. 6. 6. The soluble fraction containing 80% of ALP activity was unable to support calcium deposition. The mixing of the soluble and insoluble fractions after PI-PLC treatment failed to fully restore calcium-depositing activity. |
| Databáze: | OpenAIRE |
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