Ricchelli F, Barbato P, Milani M, Gobbo S, Salet C, Moreno G
Consiglio Nazionale delle Ricerche, Centro Metalloproteine, Dipartimento di Biologia, Università di Padova, Italy.
Biochem J. 1999 Feb 15;338 ( Pt 1)(Pt 1):221-7.
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.
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。