Isolation of stable pancreatic zymogen granules.
Isolation of stable pancreatic zymogen granules.
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稳定的胰腺酶原颗粒的分离。
DOI：
10.1152/ajpgi.1984.246.4.g411
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Hopfer,U
中科院分区：
文献类型：
--
作者：
DeLisle,RC;Schulz,I;Tyrakowski,T;Haase,W;Hopfer,U
据报道，分离的胰腺酶原颗粒在常见的电解质溶液（例如 NaCl 或 KCl）或 pH 值高于 5.5 的情况下会裂解。开发了一种基于等渗 Percoll 密度梯度的新方法，用于分离酶原颗粒，并将其应用于大鼠胰腺。根据电子显微镜外观和比淀粉酶活性判断，颗粒是高度纯化的。这些颗粒在生理 pH 值和等渗 NaCl 或 KCl 中表现出高度稳定性。用库尔特计数器测定的酶原颗粒直径在等渗 NaCl 和 KCl 中均为 1.0 +/- 0.2 微米。在生理溶液中获得的这些尺寸值与通过显微镜在完整细胞中测定的颗粒尺寸相当。淀粉酶活性平均每颗粒 0.66 microU，蛋白质含量平均每颗粒 0.31 pg；这些值并没有受到 pH 值在 5.5 到 7.0 之间以及离子强度在接近 0 到 0.15 之间的不同条件的显着影响。根据蛋白质含量估算的颗粒密度为 1.13 g/ml，这与颗粒在密度梯度中的行为非常吻合。新制剂中酶原颗粒的特性纠正了它们作为储存细胞器的作用与之前报道的体外不稳定性之间的明显差异。
Isolated pancreatic zymogen granules have been reported to lyse in common electrolyte solutions such as NaCl or KCl or at pH values above 5.5. A new method, based on an isosmotic Percoll density gradient, was developed for the isolation of zymogen granules and applied to rat pancreas. The granules are highly purified as judged by electron microscopic appearance and specific amylase activity. These granules exhibit a high degree of stability at physiological pH and in isotonic NaCl or KCl. Zymogen granule diameters, determined with a Coulter Counter, were 1.0 +/- 0.2 micron in either isotonic NaCl and KCl. These size values, obtained in physiological solutions, are comparable with granule sizes determined in intact cells by microscopy. Amylase activity averaged 0.66 microU per granule and protein content averaged 0.31 pg per granule; these values were not significantly influenced by different conditions of pH between 5.5 and 7.0 and ionic strength from near 0 to 0.15. The granule density estimated from the protein content was 1.13 g/ml, which agrees well with the behavior of granules in a density gradient. The properties of zymogen granules from the new preparation rectify the apparent discrepancy between their role as a storage organelle and their previously reported in vitro instability.