Reiss O K
J Cell Biol. 1966 Jul;30(1):45-57. doi: 10.1083/jcb.30.1.45.
The isolation and partial characterization of subcellular particles from rabbit and rat lung are described. Detailed methods for separating a purified, active mitochondrial fraction are outlined and evaluated in terms of enzymatic, chemical, and morphological criteria. Mitochondrial preparations from rabbit and rat liver were used as comparative indices. The lung mitochondrial fraction was identified by its ability to oxidize succinate with a P/O ratio of 1.7 by a process sensitive to 2,4 dinitrophenol and antimycin A. The adenosine triphosphatase activity of the lung mitochondrial fraction is stimulated by magnesium ions, but this stimulation is not augmented by 2,4 dinitrophenol. In the absence of magnesium ions, the specific activity of the adenosine triphosphatase increases with increasing protein concentration. The presence of lysosomes in the mitochondrial fraction is suggested by acid phosphatase and cathepsin activities and by electron microscope observations.
本文描述了从兔和大鼠肺中分离亚细胞颗粒并进行部分特性鉴定的过程。概述了分离纯化活性线粒体组分的详细方法,并根据酶学、化学和形态学标准进行了评估。使用兔和大鼠肝脏的线粒体提取物作为比较指标。通过对2,4-二硝基苯酚和抗霉素A敏感的过程,以1.7的P/O比值氧化琥珀酸的能力来鉴定肺线粒体组分。肺线粒体组分的腺苷三磷酸酶活性受镁离子刺激,但2,4-二硝基苯酚不会增强这种刺激。在没有镁离子的情况下,腺苷三磷酸酶的比活性随蛋白质浓度的增加而增加。酸性磷酸酶和组织蛋白酶活性以及电子显微镜观察结果表明线粒体组分中存在溶酶体。