鸡骨骺软骨中的基质小泡。与溶酶体的分离及无机焦磷酸酶活性的分布

Matrix vesicles in chicken epiphyseal cartilage. Separation from lysosomes and the distribution of inorganic pyrophosphatase activity.

作者信息

Väänänen H K, Korhonen L K

出版信息

Calcif Tissue Int. 1979 Aug 24;28(1):65-72. doi: 10.1007/BF02441219.

Abstract

The extracellular matrix vesicles from epiphyseal cartilage of chickens were isolated by differential centrifugation. The matrix vesicles obtained showed considerable activity of lysosomal enzymes. This appears to have been due to lysosomal contamination because when we used a new density gradient medium (Percoll), the lysosomal enzyme activities and the activity of alkaline phosphatase could be totally separated. Electron microscopy of the alkaline phosphatase-rich fraction showed matrix vesicle-like structures. Phosphatase activities of the cells and matrix vesicles were further studied by Sephadex G-200 gel filtration. Specific magnesium-activated inorganic pyrophosphatase, distinct from nonspecific alkaline phosphatase, could be demonstrated in the cellular fraction. No such separate activity could be demonstrated in the matrix vesicle fraction, and it is supposed that the pyrophosphatase activity in the matrix vesicles originates from the alkaline phosphate.

摘要

通过差速离心法从鸡骨骺软骨中分离出细胞外基质囊泡。所获得的基质囊泡显示出相当高的溶酶体酶活性。这似乎是由于溶酶体污染所致，因为当我们使用一种新的密度梯度介质（Percoll）时，溶酶体酶活性和碱性磷酸酶活性能够完全分离。对富含碱性磷酸酶的部分进行电子显微镜观察，发现了类似基质囊泡的结构。通过Sephadex G - 200凝胶过滤进一步研究了细胞和基质囊泡的磷酸酶活性。在细胞部分可以证明存在与非特异性碱性磷酸酶不同的特异性镁激活无机焦磷酸酶。在基质囊泡部分未发现这种单独的活性，推测基质囊泡中的焦磷酸酶活性源自碱性磷酸酶。