Bellomo G, Vairetti M, Stivala L, Mirabelli F, Richelmi P, Orrenius S
Dipartimento di Medicina Interna e Terapia Medica, University of Pavia, Italy.
Proc Natl Acad Sci U S A. 1992 May 15;89(10):4412-6. doi: 10.1073/pnas.89.10.4412.
The intracellular distribution of glutathione (GSH) in cultured hepatocytes has been investigated by using the compound monochlorobimane (BmCl), which interacts specifically with GSH to form a highly fluorescent adduct. Image analysis of BmCl-labeled hepatocytes predominantly localized the fluorescence in the nucleus; the nuclear/cytoplasmic concentration gradient was approximately three. This concentration gradient was collapsed by treatment of the cells with ATP-depleting agents. The uneven distribution of BmCl fluorescence was not attributable to (i) nonspecific interaction of BmCl with protein sulfhydryl groups, (ii) any selective nuclear localization of the GSH transferase(s) catalyzing formation of the GSH-BmCl conjugate, or (iii) any apparent alterations in cell morphology from culture conditions, suggesting that this distribution did, indeed, reflect a nuclear compartmentalization of GSH. That the nuclear pool of GSH was found more resistant to depletion by several agents than the cytoplasmic pool supports the assumption that GSH is essential in protecting DNA and other nuclear structures from chemical injury.
通过使用化合物单氯双硫腙（BmCl），研究了培养的肝细胞中谷胱甘肽（GSH）的细胞内分布，该化合物与GSH特异性相互作用形成高度荧光加合物。对BmCl标记的肝细胞进行图像分析，结果显示荧光主要定位于细胞核；核/细胞质浓度梯度约为3。用ATP消耗剂处理细胞后，这种浓度梯度消失。BmCl荧光的不均匀分布并非归因于：（i）BmCl与蛋白质巯基的非特异性相互作用；（ii）催化GSH-BmCl共轭物形成的谷胱甘肽转移酶的任何选择性核定位；或（iii）培养条件导致的细胞形态的任何明显改变，这表明这种分布确实反映了GSH的核区室化。发现GSH的核库比细胞质库对几种试剂的消耗更具抗性，这支持了GSH对于保护DNA和其他核结构免受化学损伤至关重要的假设。
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