Department of Pharmacology, University of Miami School of Medicine, Biscayne Annex, P. O. Box 875, 33152, Miami, Fla..
J Membr Biol. 1972 Dec;7(1):345-64. doi: 10.1007/BF01867925.
The use of the fluorescent chelate probe, chlorotetracycline, in mitochondria is described. The probe shows a high fluorescence in the presence of mitochondria which may be ascribed to binding of the probe to membrane-associated Ca(++) and Mg(++). The fluorescence excitation and emission spectra are diagnostic of binding of the probe to Ca(++) in coupled mitochondria and Mg(++) in uncoupled mitochondria. The fluorescence polarization spectra are diagnostic of the cations having a moderately high mobility in the membrane environment. The effects of exogenous EDTA and of endogenous Mn(++) indicate that the probe is primarily visualizing actively accumulated Ca(++) on the inner surface of the inner membrane. By employing the Ca(++) transport inhibitor, Tb(+++), the fluorescence changes associated with metabolic alterations are shown to arise partly from cation transport and partly through alterations in the binding properties of the inner surface of the membrane. Chlorotetracycline is a probe for divalent cations associated with the membrane and is of general utility in the study of cation migrations in cellular and subcellular systems.
本文描述了荧光螯合物探针——金霉素在线粒体中的应用。该探针在存在线粒体的情况下显示出高荧光，这可能归因于探针与膜相关的 Ca(++)和 Mg(++)结合。荧光激发和发射光谱是探针与耦联线粒体中的 Ca(++)和去耦联线粒体中的 Mg(++)结合的诊断指标。荧光偏振光谱是膜环境中具有中等高迁移率的阳离子的诊断指标。外源性 EDTA 和内源性 Mn(++)的影响表明，探针主要是可视化主动积累的 Ca(++)在内膜内表面上。通过使用 Ca(++)转运抑制剂 Tb(+++)，显示与代谢变化相关的荧光变化部分来自阳离子转运，部分来自膜内表面的结合特性的变化。金霉素是一种与膜相关的二价阳离子探针，在研究细胞和亚细胞系统中的阳离子迁移方面具有普遍的用途。
