溶酶体酶在嗜铬组织中的定位。

The localization of lysosomal enzymes in chromaffin tissue.

作者信息

Smith A D, Winkler H

出版信息

J Physiol. 1966 Mar;183(1):179-88. doi: 10.1113/jphysiol.1966.sp007859.

DOI:10.1113/jphysiol.1966.sp007859

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1357535/

Abstract

An homogenate of bovine adrenal medulla contains significant amounts of six acid hydrolases: acid ribonuclease, acid deoxyribonuclease, cathepsin, acid phosphatase, beta-glucuronidase and arylsulphatase. Most of the activity of each enzyme could be sedimented in the large-granule fraction at 242,000 g-min.2. Differential centrifugation indicated the presence of three populations of particles, which sedimented at slightly different rates; these are, in order of decreasing sedimentation rate, mitochondria, particles containing the acid hydrolases, and chromaffin granules.3. The three types of particle could be separated by ultracentrifuging the large-granule fraction in a sucrose density gradient. Most of the activity of each hydrolase was recovered in a layer intermediate between those formed by mitochondria and chromaffin granules.4. The large-granule fraction therefore contains particles which are defined by their enzyme content as lysosomes.5. Highly purified chromaffin granules, containing less than 5% of the activity of each acid hydrolase, were obtained from the gradient.

摘要

牛肾上腺髓质匀浆含有大量六种酸性水解酶：酸性核糖核酸酶、酸性脱氧核糖核酸酶、组织蛋白酶、酸性磷酸酶、β-葡萄糖醛酸酶和芳基硫酸酯酶。每种酶的大部分活性可在242,000克离心分钟的大颗粒组分中沉降。
差速离心表明存在三类颗粒，它们以略有不同的速率沉降；按沉降速率递减顺序依次为线粒体、含有酸性水解酶的颗粒和嗜铬颗粒。
通过在蔗糖密度梯度中对大颗粒组分进行超速离心可分离出这三种类型的颗粒。每种水解酶的大部分活性在由线粒体和嗜铬颗粒形成的层之间的中间层中回收。
因此，大颗粒组分含有因其酶含量而被定义为溶酶体的颗粒。
从梯度中获得了高度纯化的嗜铬颗粒，其每种酸性水解酶的活性含量低于5%。