LYSOSOMES OF ARTERIAL WALL .1. ISOLATION AND SUBCELLULAR FRACTIONATION OF CELLS FROM NORMAL RABBIT AORTA
LYSOSOMES OF ARTERIAL WALL .1. ISOLATION AND SUBCELLULAR FRACTIONATION OF CELLS FROM NORMAL RABBIT AORTA
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DOI：
10.1084/jem.136.5.1117
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发表时间：
1972-01-01
影响因子：
15.3
通讯作者：
MULLER, M
中科院分区：
文献类型：
--
作者：
PETERS, TJ;DEDUVE, C;MULLER, M
用含弹性蛋白酶、胶原酶和透明质酸酶的Hanks‘s液孵育正常兔主动脉，分离出正常兔主动脉的平滑肌细胞。分离得到的细胞含有大量的酸性水解酶：N-乙酰-β-氨基葡萄糖苷酶、N-乙酰-β-半乳糖苷酶、β-半乳糖苷酶、β-葡萄糖苷酶、α-甘露糖苷酶、β-葡萄糖苷酶、酸性磷酸酶、组织蛋白酶C和D。在这些蔗糖梯度中研究的其他颗粒酶包括细胞色素氧化酶和单胺氧化酶(线粒体)、5‘-核苷酸酶和亮氨酰-β-萘酰胺酶(质膜)和过氧化氢酶(？这种微量分析亚细胞分离技术适用于许多其他组织的毫克量的研究，包括正常组织和病理组织。
Smooth muscle cells were dissociated from normal rabbit aorta by incubating the tissue in Hanks' solution containing elastase, collagenase, and hyaluronidase. The isolated cells contained significant amounts of the following acid hydrolases:N-acetyl-ß-glucosaminidase,N-acetyl-ß-galactosaminidase, ß-galactosidase, ß-glucuronidase, α-mannosidase, ß-glucosidase, acid phosphatase, and cathepsins C and D. The cells were disrupted and fractionated by isopycnic centrifugation on sucrose density gradients in the Beaufay automatic zonal rotor.Lysosomes with a modal density of 1.16 were identified by the distribution of these acid hydrolases and by the latency ofN-acetyl-ß-glucosaminidase and ß-galactosidase. Other particulate enzymes studied in these sucrose gradients included cytochrome oxidase and monoamine oxidase (mitochondria), 5'-nucleotidase and leucyl-ß-naphthylamidase (plasma membrane), and catalase (? peroxisome).This microanalytical subcellular fractionation technique is applicable to the study of milligram quantities of many other tissues, both normal and pathological.