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膜硫醇基团光氧化诱导溶酶体渗透敏感性增强。

Enhancement of lysosomal osmotic sensitivity induced by the photooxidation of membrane thiol groups.

作者信息

Wan Feng-Yi, Yang Lu, Zhong Yi-Gang, Zhu Wen, Wang Yi-Nan, Zhang Guo-Jiang

机构信息

Department of Cellular Biophysics, Institute of Biophysics, Academia Sinica, Beijing, People's Republic of China.

出版信息

Photochem Photobiol. 2002 Feb;75(2):134-9. doi: 10.1562/0031-8655(2002)075<0134:eolosi>2.0.co;2.

Abstract

The osmotic lysis of photodamaged lysosomes is a critical event for killing tumor cells. How the photodamage increases lysosomal osmotic sensitivity is still unclear. In this work, the effect of the photooxidation of membrane thiol groups on the lysosomal osmotic sensitivity was studied by measuring the thiol groups with 5,5'-dithiobis(2-nitrobenzoic acid) and examining the lysosomal beta-hexosaminidase latency loss in a hypotonic sucrose medium. The results show that methylene blue-mediated photooxidation of lysosomes decreased their membrane thiol groups and produced cross-linkage of membrane proteins (molecular weight ranging from 75000 to 125000), which was visualized by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Simultaneously, the lysosomal osmotic sensitivity increased. These photoinduced alterations of the lysosomes could be recovered by reducing the oxidized thiol groups with dithiothreitol. It indicates that the photooxidation of membrane thiol groups can increase the lysosomal osmotic sensitivity and therefore provides a new explanation for the photoinduced lysosomal lysis.

摘要

光损伤溶酶体的渗透性裂解是杀死肿瘤细胞的关键事件。光损伤如何增加溶酶体的渗透敏感性仍不清楚。在这项工作中,通过用5,5'-二硫代双(2-硝基苯甲酸)测量巯基并检测低渗蔗糖培养基中溶酶体β-己糖胺酶潜伏性丧失,研究了膜巯基的光氧化对溶酶体渗透敏感性的影响。结果表明,亚甲蓝介导的溶酶体光氧化降低了其膜巯基,并产生了膜蛋白的交联(分子量范围为75000至125000),这通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳得以可视化。同时,溶酶体的渗透敏感性增加。这些溶酶体的光诱导改变可以通过用二硫苏糖醇还原氧化的巯基来恢复。这表明膜巯基的光氧化可以增加溶酶体的渗透敏感性,因此为光诱导的溶酶体裂解提供了新的解释。

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