PREPARATION AND PROPERTIES OF PLASMA MEMBRANE AND ENDOPLASMIC RETICULUM FRAGMENTS FROM ISOLATED RAT FAT CELLS
PREPARATION AND PROPERTIES OF PLASMA MEMBRANE AND ENDOPLASMIC RETICULUM FRAGMENTS FROM ISOLATED RAT FAT CELLS
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DOI：
10.1016/0005-2736(71)90331-2
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发表时间：
1971-01-01
期刊：
影响因子：
--
通讯作者：
WALLACH, DFH
中科院分区：
文献类型：
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作者：
AVRUCH, J;WALLACH, DFH
1 .采用蔗糖梯度离心分离法，将游离脂肪细胞的质膜和内质网作为混合微粒体分离，获得高产率。2. 通过观察连续聚合物（葡聚糖）梯度在一定pH值和离子条件下等渗密度的变化，探索了这些颗粒的半透性行为。使用不连续的葡聚糖梯度，质膜部分纯化。质膜组分显示在匀浆上富集了2- 16倍的5′-核苷酸酶。Na+-K+刺激的atp酶在这部分中以最高的比活性恢复。线粒体污染为3%，内质网污染为2.7%。描述了一种灵敏的5′-核苷酸酶放射测定法。
I. Plasma membrane and endoplasmic reticulum of free fat cells were isolated in high yield as a mixed microsomal fraction by sucrose gradient centrifugation. 2. The semipermeable behavior of these particles was explored by observing shifts in isopycnic density in continuous polymer (Dextran) gradients over a range of pH and ionic conditions.3. Using a discontinuous Dextran gradient, a plasma membrane fraction was partially purified. The plasma membrane fraction showed a I2-i6-fold enrichment of 5'-nucleotidase over the homogenate. Na+-K+-stimulated ATPase was recovered in highest specific activity in this fraction. Mitochondrial contamination was 3% where endoplasmic reticulum contamination was 2 7%. 4-A sensitive radioassay for 5'-nucleotidase is described.