Oxidative phosphorylation and ultrastructural transformation in mitochondria in the intact ascites tumor cell.
Oxidative phosphorylation and ultrastructural transformation in mitochondria in the intact ascites tumor cell.
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DOI：
10.1083/jcb.51.1.123
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发表时间：
1971-10
期刊：
影响因子：
--
通讯作者：
Lemasters JJ
中科院分区：
文献类型：
--
作者：
Hackenbrock CR;Rehn TG;Weinbach EC;Lemasters JJ
我们已经检查了线粒体的超微结构，因为它与完整细胞中的能量代谢有关。在超微结构完整的艾氏腹水肿瘤细胞中，通过添加2-脱氧葡萄糖快速从内源性三磷酸腺苷生成二磷酸腺苷，诱导细胞内氧化磷酸化。通过连续同步监测呼吸频率、吡啶核苷酸荧光和90°光散射，间接确定氧化磷酸化的发生。细胞内腺嘌呤核苷酸的直接酶分析和细胞内无机正磷酸盐的测定证实了氧化磷酸化。电子显微镜快速固定的细胞显微样品显示，除氧化磷酸化外，所有细胞的线粒体在2-脱氧葡萄糖生成二磷酸腺苷后不到6s内都发生了传统的→浓缩的超微结构转变。90°光散射增加，这也发生在这个时候，显示出只有25秒的时间，这与较慢的正统→最大浓缩线粒体转化在时间上是一致的。在氧化磷酸化解偶联剂存在的情况下，细胞内腺苷二磷酸的生成既不能启动完整细胞线粒体的氧化磷酸化，也不能启动线粒体的超微结构转化。寡霉素部分和完全抑制氧化磷酸化与部分和完全抑制2-脱氧葡萄糖诱导的线粒体超微结构转化呈正相关。这些数据表明，在完整的腹水肿瘤细胞中，正统的→浓缩超微结构转化与诱导线粒体氧化磷酸化有关。
We have examined the ultrastructure of mitochondria as it relates to energy metabolism in the intact cell. Oxidative phosphorylation was induced in ultrastructurally intact Ehrlich ascites tumor cells by rapidly generating intracellular adenosine diphosphate from endogenous adenosine triphosphate by the addition of 2-deoxyglucose. The occurrence of oxidative phosphorylation was ascertained indirectly by continuous and synchronous monitoring of respiratory rate, fluorescence of pyridine nucleotide, and 90° light-scattering. Oxidative phosphorylation was confirmed by direct enzymatic analysis of intracellular adenine nucleotides and by determination of intracellular inorganic orthophosphate. Microsamples of cells rapidly fixed for electron microscopy revealed that, in addition to oxidative phosphorylation, an orthodox → condensed ultrastructural transformation occurred in the mitochondria of all cells in less than 6 sec after the generation of adenosine diphosphate by 2-deoxyglucose. A 90° light-scattering increase, which also occurs at this time, showed a t ½ of only 25 sec which agreed temporally with a slower orthodox → maximally condensed mitochondrial transformation. Neither oxidative phosphorylation nor ultrastructural transformation could be initiated in mitochondria in intact cells by the intracellular generation of adenosine diphosphate in the presence of uncouplers of oxidative phosphorylation. Partial and complete inhibition of oxidative phosphorylation by oligomycin resulted in a positive relationship to partial and complete inhibition of 2-deoxyglucose-induced ultrastructural transformation in the mitochondria in these cells. The data presented reveal that an orthodox → condensed ultrastructural transformation is linked to induced oxidative phosphorylation in mitochondria in the intact ascites tumor cell.