The mobility of a fluorescent ubiquinone in model lipid membranes. Relevance to mitochondrial electron transport.
The diffusion and location of a functional, fluorescent ubiquinone molecule, NBDHA-Q, were determined as a function of temperature using microscopic observation, fluorescence recovery after photobleaching and fluorescence spectroscopy in protein-free, pure-lipid dimyristoylphosphatidylcholine and dimyristoylphosphatidylcholine/cholesterol multibilayers. The data reveal that in a liquid-crystalline membrane (1) ubiquinone is highly mobile, (2) ubiquinone uniformly diffuses laterally with the same diffusion coefficient (3.10(-8) cm2/s at 25 degrees C) as the phospholipids in which it resides, (3) the diffusion coefficients of ubiquinone and phospholipid both decrease at the exothermic phase transition of the phospholipid, (4) cholesterol affects the diffusion coefficients of ubiquinone and phospholipids to the same degree, (5) cholesterol induces a lateral phase separation progressively excluding ubiquinone from cholesterol-containing domains. These data suggest that ubiquinone does not reside at the membrane surface or in the mid-plane for any appreciable length of time. Rather, the data indicate that ubiquinone is highly mobile laterally and transversely, spending the majority of its time in the acyl chain region of the membrane, where its lateral and transverse diffusion is limited by the lateral diffusion and the transverse microviscosity gradient of the phospholipids and where its lateral location can be affected by the presence of cholesterol. In addition, based upon a comparison of the diffusion coefficients for ubiquinone, phospholipids and mitochondrial redox complexes, we hypothesize that no significant portion of the ubiquinone pool remains bound to redox complexes for any significant length of time relative to that for electron transport as resolvable by fluorescence recovery after photobleaching.
Chazotte
B
B
Department of Cell Biology and Anatomy, University of North Carolina, School of Medicine, Chapel Hill 27599.
Wu
E S
ES
Hackenbrock
C R
CR
D002784:Cholesterol / Q000378:metabolism; D004058:Diffusion; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D004579:Electron Transport; D005453:Fluorescence; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism*; D050356:Lipid Metabolism; D008928:Mitochondria / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D013696:Temperature; D014451:Ubiquinone / Q000378:metabolism*; D014783:Viscosity
D008051:Lipid Bilayers; D010743:Phospholipids; D014451:Ubiquinone; D002784:Cholesterol; D004134:Dimyristoylphosphatidylcholine