Nilsson K B, Andersson R G, Eneström S, Mackerlova L, Mohme-Lundholm E
Acta Pharmacol Toxicol (Copenh). 1978 Mar;42(3):194-205. doi: 10.1111/j.1600-0773.1978.tb02190.x.
From a homogenate of rabbit colon muscle subcellular fractions were isolated by differential centrifugation. The crude microsomal fraction could be separated into subfractions, a fraction of vesicular microsomes at 35% sucrose, a fraction containing sarcolemma, mitochondrial fragments and microsomal vesicles at 35--45% sucrose and a small protein fraction at 45--55% sucrose. Their biochemical properties and their morphological characterization were investigated. The cholesterol and the phospholipid content was equally distributed between the microsomal fractions 35% and 35--45% while the RNA was localized to the mitochondria and the microsomal fraction 35%. The enzyme cytochrome c oxidase was found to be concentrated in the mitochondria while a high contamination was found in the microsomal fractions 35--45%. The NADH-oxidase activity was highest in the 35% fraction and the 5'-nucleotidase activity in the 40,000 X g supernatant. The microsomal subfractions contained the enzymes ATPase, adenylate cyclase and phosphodiesterase. In the 35% fraction Ca stimulated the hydrolysis of ATP. The binding of [3H]-ouabain and the incorporation of [3H]-leucine was most pronounced in the 35% fraction. In a K+-free Krebs Ringer medium the binding of the glucoside was stimulated in all the fractions. From these results we concluded that the fraction 35% sucrose may be mainly derived from the endoplasmic reticulum and the plasma membrane while the 35--45% originates from the plasma membrane, mitochondria and to a lesser extent the endoplasmic reticulum.
通过差速离心从兔结肠肌肉匀浆中分离出亚细胞组分。粗微粒体组分可进一步分离成亚组分，即在35%蔗糖浓度下的囊泡微粒体组分、在35% - 45%蔗糖浓度下包含肌膜、线粒体片段和微粒体囊泡的组分以及在45% - 55%蔗糖浓度下的小蛋白质组分。对它们的生化特性和形态特征进行了研究。胆固醇和磷脂含量在35%和35% - 45%的微粒体组分中分布均匀，而RNA定位于线粒体和35%的微粒体组分中。发现细胞色素c氧化酶集中在线粒体中，而在35% - 45%的微粒体组分中发现有高污染。NADH氧化酶活性在35%的组分中最高，5'-核苷酸酶活性在40,000×g上清液中最高。微粒体亚组分含有ATP酶、腺苷酸环化酶和磷酸二酯酶。在35%的组分中，Ca刺激ATP的水解。[3H]-哇巴因的结合和[3H]-亮氨酸的掺入在35%的组分中最为明显。在无K+的 Krebs Ringer培养基中，所有组分中糖苷的结合均受到刺激。根据这些结果，我们得出结论，35%蔗糖浓度的组分可能主要来源于内质网和质膜，而35% - 45%的组分则来源于质膜、线粒体，在较小程度上来源于内质网。
