Mobility of fluorescent derivatives of cytochrome c in mitochondria.
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.
Vanderkooi
J M
JM
Maniara
G
G
Erecinska
M
M
D000818:Animals; D003574:Cytochrome c Group / Q000378:metabolism*; D007313:Insecta; D007425:Intracellular Membranes / Q000502:physiology; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000502:physiology; D008856:Microscopy, Fluorescence; D008931:Mitochondria, Muscle / Q000502:physiology*; D009038:Motion; D010101:Oxygen Consumption; D011166:Porphyrins
D003574:Cytochrome c Group; D008563:Membrane Lipids; D011166:Porphyrins