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卟啉对内质网中钙离子内流的光动力作用:与线粒体的比较。

Photodynamic action of porphyrin on Ca2+ influx in endoplasmic reticulum: a comparison with mitochondria.

作者信息

Ricchelli F, Barbato P, Milani M, Gobbo S, Salet C, Moreno G

机构信息

Consiglio Nazionale delle Ricerche, Centro Metalloproteine, Dipartimento di Biologia, Università di Padova, Italy.

出版信息

Biochem J. 1999 Feb 15;338 ( Pt 1)(Pt 1):221-7.

Abstract

We have studied the distribution properties of haematoporphyrin (HP) and protoporphyrin (PP) in mitochondria and endoplasmic reticulum after isolation from rat liver. The photosensitizing efficiency of porphyrin on the Ca2+ influx function of microsomes has been compared with that obtained on Ca2+ uptake in mitochondria. HP and PP are accumulated in microsomes to a greater extent than in mitochondria, both porphyrins binding to membrane protein sites. The Ca2+ influx functions of mitochondria and microsomes, before and after irradiation in the presence of HP or PP, were studied by following the changes in the free Ca2+ concentration in the medium as revealed by the variations in fluorescence intensity of the Ca2+ indicator Calcium Green-1. For the same amount of incorporated porphyrin, the Ca2+ influx function of microsomes is degraded by irradiation more rapidly than that of mitochondria. The protective effect of dithiothreitol suggests that thiol groups in the Ca2+-transporting enzyme are the preferential targets of the photodynamic effect. These results suggest that intracellular Ca2+ movements are altered primarily by the endoplasmic reticulum rather than by mitochondrial damage, in good agreement with other observations made in porphyrin-loaded irradiated cells.

摘要

我们研究了从大鼠肝脏分离后,血卟啉(HP)和原卟啉(PP)在线粒体和内质网中的分布特性。将卟啉对微粒体Ca2+内流功能的光敏效率与对线粒体Ca2+摄取的光敏效率进行了比较。HP和PP在微粒体中的积累程度比在线粒体中更大,两种卟啉均与膜蛋白位点结合。通过追踪Ca2+指示剂钙黄绿素-1荧光强度的变化所揭示的培养基中游离Ca2+浓度的变化,研究了在HP或PP存在下照射前后线粒体和微粒体的Ca2+内流功能。对于相同量掺入的卟啉,微粒体的Ca2+内流功能比线粒体的Ca2+内流功能在照射后降解得更快。二硫苏糖醇的保护作用表明,Ca2+转运酶中的巯基是光动力效应的优先靶点。这些结果表明,细胞内Ca2+的移动主要是由内质网改变的,而不是由线粒体损伤改变的,这与在卟啉负载的照射细胞中进行的其他观察结果一致。

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