Biochemical studies of rat liver Golgi apparatus. I. Isolation and preliminary characterization.
Biochemical studies of rat liver Golgi apparatus. I. Isolation and preliminary characterization.
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大鼠肝脏高尔基体的生化研究。
DOI：
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发表时间：
1978
期刊：
影响因子：
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通讯作者：
S. Shimizu
中科院分区：
文献类型：
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作者：
Y. Hino;A. Asano;R. Sato;S. Shimizu
本文报道了一种从大鼠肝脏中分离形态学保存完好的高尔基体的方法。该方法基本上与Morré等人（Morré，D. J.，汉密尔顿，R.L.，Mollenhauser，H.H.，Mahley，R.W.，坎宁安，W. P.，Cheetham，R. D.，& Lequire，V.S.（1970）J. Cell Biol.44，484-491），不同之处在于通过不锈钢筛实现温和的细胞破碎。蛋白质和半乳糖基转移酶的平均回收率为约6毫克，从10克灌注的肝脏和约35%的匀浆，分别。该制剂实际上不含琥珀酸-细胞色素c还原酶、葡萄糖-6-磷酸酶、酸性磷酸酶和5 '-核苷酸酶。在蔗糖和葡聚糖密度梯度中等密度离心后，高尔基体部分以及其囊泡片段是均匀的。它们在蔗糖中的浮力密度明显高于在葡聚糖中的浮力密度，表明这两种形式的细胞器都是封闭的系统，对大分子是不可渗透的。半乳糖基转移酶活性的新鲜制备的高尔基体部分，测量与卵清蛋白作为半乳糖基受体，被激活26倍，通过添加Triton X-100，而那些均质化，超声处理，和老化的制剂仅激活2至4倍。
A method is described for the isolation of morphologically well-preserved Golgi apparatus from rat liver. The method is essentially the same as that of Morré et al. (Morré, D.J., Hamilton, R.L., Mollenhauser, H.H., Mahley, R.W., Cunningham, W.P., Cheetham, R.D., & Lequire, V.S. (1970) J. Cell Biol. 44, 484-491) except that mild cell disruption is achieved by means of a stainless-steel sieve. The average recoveries of protein and galactosyltransferase in the isolated fraction are about 6 mg from 10 g of perfused liver and about 35% from the homogenate, respectively. The preparation is virtually free from succinate-cytochrome c reductase, glucose-6-phosphatase, acid phosphatase, and 5'-nucleotidase. The Golgi fraction as well as its vesicular fragments is homogeneous upon isopycnic centrifugation in both sucrose and dextran density gradients. Their buoyant densities in sucrose are significantly higher than those in dextran, indicating that both forms of the organelle are closed systems which are impermeable to macromolecules. The galactosyltransferase activity of a freshly prepared Golgi fraction, measured with ovalbumin as galactosyl acceptor, is activated 26-fold by the addition of Triton X-100, whereas those of homogenized, sonicated, and aged preparations are only activated 2- to 4-fold.