Microdynamics of outer and inner membranes of mitochondria from bullfrog myocardium.
Outer and inner mitochondrial membranes were separated from bullfrog myocardium. Membrane viscosity and wobbling angle of phospholipids were measured with a nanosecond time-resolved fluorometer using a fluorophore, DPH, in each membrane. Measurements were also made on liposomes prepared from lipids extracted from each membrane. The anisotropy decay curve for DPH fluorescence was assumed to represent a mean value of decays in several microenvironments in membranes. Phospholipid constituents in membranes were analyzed by HPLC. A high proportion of PE and CL, both of which contain large amounts of unsaturated acyl chain, were found in the inner membrane. The low viscosity and large wobbling angle of phospholipids in the liposome from the inner membrane were consistent with the probable high content of unsaturated acyl chains and the low content of cholesterol in the inner membrane. Measurements of the dynamic microstructure of mitochondrial membranes suggested multifactorial characteristics probably resulting from the lipid-protein interactions. The average viscosity was found to be 0.39 +/- 0.08 P in the outer membrane and 0.58 +/- 0.01 P in the inner membrane. The wobbling angle of phospholipids in the outer and the inner membrane was, respectively, 47 and 49 degrees (non-significant difference). The liposomes prepared from lipid extracts of the membranes showed a lower viscosity and/or higher wobbling angle of phospholipids compared with the membranes themselves. The difference in the viscosity and wobbling angle between the mitochondrial membrane and its respective liposome was large in the inner membrane. These results suggest that the motion of phospholipids is limited by membrane proteins and the limitation of molecular motion of phospholipids results in an increase in the average viscosity. The results also suggest that the microdynamic values obtained in the inner membrane fraction reflect the interaction between phospholipid and protein which is abundant in the inner membrane.
Koyama
T
T
Section of Physiology, Hokkaido University, Sapporo, Japan.
Zhu
M Y
MY
Araiso
T
T
Kinjo
M
M
D000818:Animals; D002308:Cardiolipins / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D005470:Fluorometry; D007425:Intracellular Membranes / Q000502:physiology*; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D008929:Mitochondria, Heart / Q000737:chemistry*; D008995:Monoamine Oxidase / Q000032:analysis; D010743:Phospholipids / Q000032:analysis; D011892:Rana catesbeiana; D013109:Sphingomyelins / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000032:analysis
D002308:Cardiolipins; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D013109:Sphingomyelins; D013385:Succinate Dehydrogenase; D008995:Monoamine Oxidase