P-31 NUCLEAR MAGNETIC-RESONANCE STUDIES OF ISOLATED RAT-LIVER CELLS
P-31 NUCLEAR MAGNETIC-RESONANCE STUDIES OF ISOLATED RAT-LIVER CELLS
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DOI：
10.1038/273554a0
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发表时间：
1978-01-01
期刊：
影响因子：
64.8
通讯作者：
WILLIAMSON, JR
中科院分区：
文献类型：
--
作者：
COHEN, SM;OGAWA, S;WILLIAMSON, JR
近年来，核磁共振技术已被用于完整细胞的研究，并观察和鉴定了许多磷代谢物。这些研究的一个有价值的结果是利用无机磷(PI)的31P核磁共振峰的位置随着PII质子化而移动的事实来确定细胞内的pH(参考文献1-3)。对通电的大肠杆菌3的31P核磁共振测量表明，跨膜的δpH与以前的δpH测量结果一致，该测量是由内外体积之间的弱酸(如DMO)的分布确定的4。虽然从分离的线粒体5-8的研究中有相当多的证据表明APH值的存在，但还没有报道直接测量整个细胞中胞浆和线粒体之间的δ差异。然而，从对受损大鼠肝细胞胞浆和线粒体内容物之间阴离子分布的研究中，有间接证据表明存在这种梯度。在这份初步报告中，我们证明了31P核磁共振可以用来检测大鼠肝细胞中的磷代谢物，并直接测量完整细胞中线粒体和胞浆之间的δpH。
PHOSPHOROUS nuclear magnetic resonance (NMR) has recently been used in studies of intact cells, and many phosphate metabolites have been observed and identified. One valuable result of these studies has been the determination of intracellularpH (refs 1–3) using the fact that the position of the31P NMR peak of inorganic phosphate (Pi) shifts as Piis protonated.31P NMR measurements of energisedEscherichia coli3have shown that a δpH is maintained across the membranes in agreement with previous measurements of δpH determined from the distribution of a weak acid, such as DMO, between the inner and outer volumes4. Although there is considerable evidence for the existence of δpH from studies of isolated mitochondria5–8, no direct measurements have been reported of apH difference between cytosol and mitochondria in whole cellsin vivo. However, there is indirect evidence for such a gradient from studies of the distribution of anions between the cytoplasmic and mitochondrial contents of disrupted rat liver cells9. In this preliminary report we show that31P NMR can be used to detect phosphate metabolites in rat liver cells and to measure directly δpH between mitochondria and cytosol in the intact cell.