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大鼠基质小泡的体外钙沉积：碱性磷酸酶与膜的结合对于基质小泡介导的钙沉积是否至关重要？

In vitro Ca deposition by rat matrix vesicles: is the membrane association of alkaline phosphatase essential for matrix vesicle-mediated calcium deposition?

作者信息

Hsu H H, Morris D C, Davis L, Moylan P, Anderson C H

机构信息

Department of Pathology and Oncology, University of Kansas Medical Center, Kansas City 66160-7410.

出版信息

Int J Biochem. 1993 Dec;25(12):1737-42. doi: 10.1016/0020-711x(88)90301-1.

DOI:10.1016/0020-711x(88)90301-1
PMID:8138010
Abstract
  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. About 20% hydrolytic activity was not released from MV membranes by PI-PLC treatment. 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. The specific activity of the released ALP was at least 5-fold higher than the residual activity. 5. After PI-PLC treatment, MVs also demonstrated an 80% reduction of AMP- or beta GP-dependent calcium deposition. 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.
摘要
  1. 用磷脂酰肌醇磷脂酶C（PI-PLC）处理佝偻病大鼠的基质小泡（MVs），可使约80%与膜结合的碱性磷酸酶（ALP）、AMP酶、焦磷酸酶释放到培养基中。2. 约20%的水解活性经PI-PLC处理后未从MV膜上释放出来。3. SDS-聚丙烯酰胺凝胶电泳和蛋白质印迹分析表明，PI-PLC处理后的可溶性部分中，仅有一种与ALP分子量相对应的免疫反应性蛋白。4. 释放的ALP的比活性至少比残留活性高5倍。5. 经PI-PLC处理后，MVs还表现出依赖AMP或β-GP的钙沉积减少80%。6. 含有80%ALP活性的可溶性部分无法支持钙沉积。PI-PLC处理后的可溶性部分和不溶性部分混合后，未能完全恢复钙沉积活性。

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