Barrow P C, Holt S J
Biochem J. 1971 Nov;125(2):545-55. doi: 10.1042/bj1250545.
The distribution of esterase in subcellular fractions of rat liver homogenates was compared with that of the lysosomal enzyme acid phosphatase and the microsomal enzyme glucose 6-phosphatase. Most of the esterase from sucrose homogenate sediments with glucose 6-phosphatase and about 8% is recovered in the supernatant. However, up to 53% of the esterase can be washed from microtome sections of unfixed liver, in which less cellular damage would be expected than that caused by homogenization. About 40% of both esterase and acid phosphatase are recovered in the soluble fraction after homogenization in aqueous glycerol or in a two-phase system (Arcton 113-0.25m-sucrose), although glucose 6-phosphatase is still recovered in the microsomal fraction of such homogenates. The esterase of the microsomal fraction prepared from a sucrose homogenate is much more readily released by treatment with 0.26% deoxycholate than are other constituents of this fraction. The release of esterase from the microsomal fraction by the detergent and its concomitant release with acid phosphatase after homogenization in glycerol or the two-phase system suggests that a greater proportion of esterase may be present in lysosomes of the intact cell than is indicated by the results of standard fractionation procedures.
将大鼠肝脏匀浆亚细胞组分中的酯酶分布与溶酶体酶酸性磷酸酶和微粒体酶葡萄糖6 -磷酸酶的分布进行了比较。蔗糖匀浆中的大部分酯酶与葡萄糖6 -磷酸酶一起沉淀，约8%的酯酶存在于上清液中。然而，在未固定肝脏的切片中，高达53%的酯酶可以被洗脱下来，预计其中细胞损伤比匀浆造成的要少。在水甘油或两相系统（Arcton 113 - 0.25m -蔗糖）中匀浆后，酯酶和酸性磷酸酶约40%存在于可溶部分，尽管葡萄糖6 -磷酸酶仍存在于此类匀浆的微粒体部分。用0.26%脱氧胆酸盐处理从蔗糖匀浆制备的微粒体部分时，其中的酯酶比该部分的其他成分更容易释放出来。在甘油或两相系统中匀浆后，去污剂使酯酶从微粒体部分释放，且酯酶与酸性磷酸酶同时释放，这表明完整细胞溶酶体中酯酶的比例可能比标准分级分离程序的结果所显示的要高。
