Plasma membranes from rat Sertoli cells: purification and properties.
Plasma membranes from rat Sertoli cells: purification and properties.
复制标题
大鼠支持细胞的质膜：纯化和特性。
DOI：
10.1095/biolreprod35.5.1235
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发表时间：
1986
影响因子：
3.6
通讯作者：
Hall,PF
中科院分区：
文献类型：
--
作者：
Kew,D;Muffly,KE;Hall,PF
A method is reported for preparing surface (plasma) membranes from rat Sertoli cells. The procedure is based upon homogenization in bypotonic buffer, extraction in a two-phase system, and sedimentation through two sucrose density gradients. The purified membranes consist of large sheets of membrane. The identity and purity of the membranes was demonstrated by (1) electron microscopy, (2) enzyme markers, and (3) functional activities associated with the membranes (binding of follicle-stimulating hormone [FSH] and production of cyclic adenosine 5′ inonophosphate fcAMP 1 . Electron microscopy showed membranes with small fragments of cytoplasm attached to the inside of the membrane sheets. Marker enzymes for plasma membrane (5′ -nucleotidase and alkaline phosphatase) showed more than 16- and 6-fold enrichment, respectively, and other enzymes showed that contamination by nuclei, mitochondria, endoplasmic reticulum, or cytosol was negligible. Binding of FSH was found to be specific, with KD1.2 nM and the equivalent of 7500 sites per cell. This binding was enriched 20-fold compared to whole homogenate. Production of cAMP by membranes was increased by addition of FSH and by forskolin to the purified membranes in vitro.
