Watson K, Bertoli E, Griffiths D E
Biochem J. 1975 Feb;146(2):401-7. doi: 10.1042/bj1460401.
The effect of temperature on the activation energies of mitochondrial enzymes of the yeast Saccharomyces cerevisiae was examined. Non-linear Arrhenius plots with discontinuities in the temperature range 14-19 degrees C and 19-22 degrees C were observed for the respiratory enzymes and mitochondrial ATPase (adenosine triphosphatase) respectively. A straight-line Arrhenius plot was observed for the matrix enzyme, malate dehydrogenase. The activation energies of the enzymes associated with succinate oxidation, namely, succinate oxidase, succinate dehydrogenase and succinate-cytochrome c oxidoreductase, were in the range 60-85kJ/mol above the transition temperature and 90-160kJ/mol below the transition temperature. In contrast, the corresponding enzymes associated with NADH oxidation showed significantly lower activation energies, 20-35kJ/mol above and 40-85kJ/mol below the transition temperature. The discontinuities in the Arrhenius plots were still observed after sonication, treatment with non-ionic detergents or freezing and thawing of the mitochondrial membranes. Discontinuities for cytochrome c oxidase activity were only observed in freshly isolated mitochondria, and no distinct breaks were observed after storage at -20 degrees C. Mitochondrial ATPase activity still showed discontinuities after sonication and freezing and thawing, but a linear plot was observed after treatment with non-ionic detergents. The results indicate that the various enzymes of the respiratory chain are located in a similar lipid macroenvironment within the mitochondrial membrane.
研究了温度对酿酒酵母线粒体酶活化能的影响。分别观察到呼吸酶和线粒体ATP酶(腺苷三磷酸酶)在14 - 19℃和19 - 22℃温度范围内的非线性阿伦尼乌斯图,且存在间断点。对于基质酶苹果酸脱氢酶,观察到的是直线型阿伦尼乌斯图。与琥珀酸氧化相关的酶,即琥珀酸氧化酶、琥珀酸脱氢酶和琥珀酸 - 细胞色素c氧化还原酶,在转变温度以上的活化能范围为60 - 85kJ/mol,在转变温度以下为90 - 160kJ/mol。相比之下,与NADH氧化相关的相应酶显示出显著更低的活化能,在转变温度以上为20 - 35kJ/mol,在转变温度以下为40 - 85kJ/mol。在对线粒体膜进行超声处理、用非离子洗涤剂处理或冻融后,仍能观察到阿伦尼乌斯图中的间断点。细胞色素c氧化酶活性的间断点仅在新鲜分离的线粒体中观察到,在 - 20℃储存后未观察到明显的断点。线粒体ATP酶活性在超声处理和冻融后仍显示间断点,但在用非离子洗涤剂处理后观察到线性图。结果表明,呼吸链的各种酶位于线粒体内膜内相似的脂质宏观环境中。