FLUOROMETRIC MEASUREMENT OF REDUCED PYRIDINE NUCLEOTIDE IN CELLULAR AND SUBCELLULAR PARTICLES
FLUOROMETRIC MEASUREMENT OF REDUCED PYRIDINE NUCLEOTIDE IN CELLULAR AND SUBCELLULAR PARTICLES
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DOI：
10.1016/0003-2697(62)90006-4
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发表时间：
1962-01-01
影响因子：
2.9
通讯作者：
ESTABROOK, RW
中科院分区：
文献类型：
--
作者：
ESTABROOK, RW
应用商业荧光计测量吡啶核苷酸还原和氧化的生物颗粒制剂，如线粒体，酵母和细菌已被描述和证明。用大鼠肝脏线粒体荧光法测定还原吡啶核苷酸的浓度变化，与分光光度法测定吡啶核苷酸动力学和化学分析法测定的吡啶核苷酸浓度变化进行了比较。这些研究表明，大鼠肝脏线粒体中吡啶核苷酸还原的荧光增强了4- 6倍，表明线粒体中存在吡啶核苷酸的三元络合。最大的荧光增强是观察线粒体的DPNH部分，在各种代谢条件下可获得高达8至12的增加。在线粒体、酵母和大肠杆菌中观察到的荧光增强程度与纯化酶制剂中DPNH络合的先前报道进行了比较。描述了线粒体浓度和温度对吡啶核苷酸荧光测定的影响。
The application of a commercial fluorometer for the measurement of pyridine nucleotide reduction and oxidation of biological particulate preparations, such as mitochondria, yeast, and bacteria has been described and demonstrated. The changes of concentration of reduced pyridine nucleotide as measured fluorometrically with rat liver mitochondria have been compared to the spectrophotometric measurement of pyridine nucleotide kinetics and to the changes in pyridine nucleotide concentration determined by chemical analyses. These studies show a fluorescence enhancement for reduced pyridine nucleotide of rat liver mitochondria of 4- to 6-fold suggesting the presence of a ternary complexing of pyridine nucleotide in mitochondria. The greatest fluorescence enhancement is observed with the DPNH fraction of mitochondria with increases as high as 8 to 12 obtainable under various metabolic conditions. The extent of fluorescence enhancement observed with mitochondria, bakers'' yeast, and E. coli is compared to previous reports on the DPNH complexing with purified enzyme preparations. The influences of mitochondrial concentration and temperature on the measurement of pyridine nucleotide fluorescence are described.