鲁米诺和腔肠素作为线粒体膜和细菌膜中超氧化物的检测探针。

Lucigenin and coelenterazine as superoxide probes in mitochondrial and bacterial membranes.

作者信息

Kervinen Marko, Pätsi Jukka, Finel Moshe, Hassinen Ilmo E

机构信息

Department of Medical Biochemistry and Molecular Biology, University of Oulu, Box 5000, FIN-90014 Oulu, Finland.

出版信息

Anal Biochem. 2004 Jan 1;324(1):45-51. doi: 10.1016/j.ab.2003.09.004.

DOI:10.1016/j.ab.2003.09.004

PMID:14654044

Abstract

The chemiluminescent superoxide indicators lucigenin and coelenterazine were compared in rat liver submitochondrial particles and cytoplasmic membranes from Paracoccus denitrificans. Qualitative monitoring is possible with both probes, but quantitative work with lucigenin is hampered by its dependence on one-electron reduction before the photon-emitting reaction. Therefore, calibration of measurements on complex I, capable of efficient lucigenin prereduction with reduced nicotinamide adenine dinucleotide, against xanthine oxidase, which in the presence of hypoxanthine is not able to reduce the probe to a significant rate compared to complex I, may give results in error by one order of magnitude. Coelenterazine, although susceptible of storage-dependent high background chemiluminescence, does not require prereduction and is thus a more reliable probe.

摘要

在来自反硝化副球菌的大鼠肝脏亚线粒体颗粒和细胞质膜中，对化学发光超氧化物指示剂光泽精和腔肠素进行了比较。两种探针都可进行定性监测，但由于光泽精在发光反应前依赖单电子还原，因此其定量工作受到阻碍。因此，用能够被还原型烟酰胺腺嘌呤二核苷酸有效预还原光泽精的复合体I进行测量，与黄嘌呤氧化酶校准，在次黄嘌呤存在的情况下，黄嘌呤氧化酶与复合体I相比不能以显著速率还原探针，这可能会导致结果出现一个数量级的误差。腔肠素虽然易受储存依赖性高背景化学发光的影响，但不需要预还原，因此是一种更可靠的探针。