Sagnella G, Price R, Peart W
Hypertension. 1980 Sep-Oct;2(5):595-603. doi: 10.1161/01.hyp.2.5.595.
The subcellular distribution and nature of rat renal renin has been investigated by means of analytical subcellular fractionation and gel filtration on Sephadex G-100. During differential centrifugation, renin activity was recovered mainly in soluble and heavy mitochondrial fractions. On sucrose gradient centrifugation in either a conventional or in a B XIV zonal rotor, renin activity equilibrated at 1.54 M sucrose and was partially resolved from marker enzymes for mitochondria (succinate dehydrogenase), lysosomes (acid phosphatase), plasma membranes (alkaline phosphatase), and peroxisomes (catalase). On gel filtration of the soluble or extracts of the renin-granular fractions on Sephadex G-100, renin activity eluted as a single peak with an apparent molecular weight (MW) of 42,000; no change in activity was found when these fractions were acidified to pH 3.0. When kidney homogenates were prepared in the presence of the proteolytic inhibitor N-ethylmaleimide (NEM, 10 mM), whereas the renin from the granular fractions displayed a MW of 44,000, that from the soluble fraction was apparently higher (69,000). Addition of NEM (10 mM) to the soluble fraction previously shown to contain only the low MW form of renin also resulted in an apparently high MW form of renin. These results indicate that rat renal renin is associated with a mechanically fragile, distinct type of subcellular organelle. Renin within this structure is of the low MW form and is not acid activatable. The soluble fraction, however, contains a factor(s) that, in the presence of NEM, combines with the low MW renin to form a complex of apparently high MW.
通过分析亚细胞分级分离和在葡聚糖凝胶G - 100上的凝胶过滤，对大鼠肾脏肾素的亚细胞分布和性质进行了研究。在差速离心过程中，肾素活性主要在可溶性和重线粒体组分中回收。在常规或B XIV区带转子中进行蔗糖梯度离心时，肾素活性在1.54 M蔗糖处达到平衡，并与线粒体（琥珀酸脱氢酶）、溶酶体（酸性磷酸酶）、质膜（碱性磷酸酶）和过氧化物酶体（过氧化氢酶）的标记酶部分分离。在葡聚糖凝胶G - 100上对肾素颗粒组分的可溶性部分或提取物进行凝胶过滤时，肾素活性以单一峰洗脱，表观分子量（MW）为42,000；当这些组分酸化至pH 3.0时，活性没有变化。当在蛋白水解抑制剂N - 乙基马来酰亚胺（NEM，10 mM）存在下制备肾脏匀浆时，颗粒组分中的肾素MW为44,000，而可溶性组分中的肾素MW明显更高（69,000）。向先前显示仅含有低MW形式肾素的可溶性组分中添加NEM（10 mM）也导致了明显高MW形式的肾素。这些结果表明，大鼠肾脏肾素与一种机械脆弱、独特类型的亚细胞器相关。该结构内的肾素为低MW形式，且不能被酸激活。然而，可溶性组分含有一种因子，在NEM存在下，该因子与低MW肾素结合形成明显高MW的复合物。
