细胞色素c荧光衍生物在线粒体中的移动性。
Mobility of fluorescent derivatives of cytochrome c in mitochondria.
作者信息
Vanderkooi J M, Maniara G, Erecinska M
出版信息
J Cell Biol. 1985 Feb;100(2):435-41. doi: 10.1083/jcb.100.2.435.
Motion of cytochrome c bound to giant (2-10-micron diam) mitochondria isolated from the waterbug Lethocerus indicus was examined using the technique of fluorescence recovery after photobleaching. Fluorescent cytochrome c was exchanged for native cytochrome c through partly damaged outer membrane. Recovery profiles were not statistically different when the fluorescence from iron-free cytochrome c or fluorescein-labeled cytochrome c was used and were essentially the same in the presence or absence of an uncoupler. In the presence of excess porphyrin cytochrome c, the apparent diffusion coefficient was 6 X 10(-11) cm2/s in 0.3 M sucrose-mannitol-EDTA and 3 X 10(-10) cm2/s in 0.10 M KCl/0.10 M sucrose. At concentrations of porphyrin cytochrome c that are stoichiometric with cytochrome c oxidase and for mitochondria in which excess cytochrome c was washed away, two components were observed in the recovery profile. The diffusion coefficient of the fast component was 1 X 10(-10) cm2/s. The second component showed no recovery during the time scale of measurement (D less than 10(-12) cm2/s). We speculate on the origin of the immobile fraction.
利用光漂白后荧光恢复技术，研究了与从印度田鳖分离出的巨型（直径2 - 10微米）线粒体结合的细胞色素c的运动。通过部分受损的外膜，将荧光细胞色素c与天然细胞色素c进行交换。当使用无铁细胞色素c或荧光素标记的细胞色素c的荧光时，恢复曲线在统计学上没有差异，并且在存在或不存在解偶联剂的情况下基本相同。在存在过量卟啉细胞色素c的情况下，在0.3 M蔗糖 - 甘露醇 - 乙二胺四乙酸中表观扩散系数为6×10⁻¹¹ cm²/s，在0.10 M氯化钾/0.10 M蔗糖中为3×10⁻¹⁰ cm²/s。在卟啉细胞色素c与细胞色素c氧化酶化学计量的浓度下，对于洗去过量细胞色素c的线粒体，在恢复曲线中观察到两个成分。快速成分的扩散系数为1×10⁻¹⁰ cm²/s。第二个成分在测量时间范围内没有恢复（扩散系数D小于10⁻¹² cm²/s）。我们推测了固定部分的来源。
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