Summary
A newly constructed cell, which allows simultaneous measurements of optical and electrical properties, was used to study bimolecular black membranes composed of beef heart mitochondrial lipids and their interaction with cytochromec.
The results show that these highly charged membranes are stable only in relatively limited ranges of boundary conditions. In 0.1n KCl their maximum direct current (dc) resistance is 7×108 Ohm cm2±10%; the series capacity at 1kHz is 0.43 μF/cm2±3%; the entire thickness, determined by optical reflectivity, is 5.8±0.2 nm.
The interaction between oxidized cytochromec and these lipid membranes is primarily of electrostatic nature, and dependent on the presence of highly charged phospholipids, such as diphosphatidyl glycerol (cardiolipin) and phosphatidyl ethanolamine. The attachment of cytochromec maximally causes a 2.5-fold increase in reflectivity, without any noticeable change in the capacity. This leads to a subsequent instability of the membrane (i.e., rupture) preceded by a rapid increase of the dc conductivity. This behavior is far less pronounced with reduced cytochromec.
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References
Cherry, R.J., Chapman, D. 1969a. Optical properties of black lecithin films.J. Mol. Biol. 40:19
Cherry, R.J., Chapman, D. 1969b. Optical determination of the thickness of thin lipid films.J. Theor. Biol. 24:137
Das, M.L., Crane, F.L. 1964. Proteolipids. I. Formation of phospholipid-cytochromec complexes.Biochemistry 3:696
Das, M.L., Haak, E.D., Crane, F.L. 1965. Proteolipids. IV. Formation of complexes between cytochromec and purified phospholipids.Biochemistry 4:859
Dawson, R.M.C., Quinn, P.J. 1971. The interaction of soluble proteins with lipid interfaces.In: Advances in Experimental Medicine and Biology. Vol.14, p. 1. Plenum Press, New York
De Vries, A.J. 1958. Foam stability. Recl. Trav. Chim. Pays-Bas. IV. Kinetics and activation energy of film rupture.77:383
Ernster, L., Kuylenstierna, B. 1970. Outer membranes of mitochondria.In: Membranes of Mitochondria and Chloroplasts. p. 172. Van Nostrand Reinhold Co., New York & London
Fleischer, S., Brierley, G., Klouwen, H., Slautterback, D.B. 1962. Studies of the electron transfer system. XLVII. The role of phospholipids in electron transfer.J. Biol. Chem. 237:3264
Fleischer, S., Rouser, G., Fleischer, B., Casu, A., Kritchevsky, G. 1967. Lipid composition of mitochondria from bovine heart, liver, and kidney.J. Lipid Res. 8:170
Folch, J., Lees, M., Stanley, G.H.S. 1957. A simple method of the isolation and purification of total lipids from animal tissues.J. Biol. Chem. 226:497
Gitler, C., Montal, M. 1972a. Thin proteolipid films: A new approach to the reconstitution of biological membranes.Biochem. Biophys. Res. Commun. 47:1486
Gitler, C., Montal, M. 1972b. Formation of decane-soluble proteolipids: Influence of monovalent and divalent cations.FEBS Lett. 28:329
Kimelberg, H.K., Lee, C.P., Claude, A., Mrena, E. 1970. Interactions of cytochromec with phospholipid membranes. I. Binding of cytochromec to phospholipid liquid crystals.J. Membrane Biol. 2:235
Margoliash, E., Schejter, A. 1966. Cytochromec Adv. Protein Chem. 21:113
Mueller, P., Rudin, C.P., Ti, Tien, H., Wescott, W.C. 1962. Reconstitution of cell membrane structure in vitro and its transformation into an excitable system.Nature 194:980
Nöll, G.G. 1976. A Cell for simultaneous measurements of optical and electrical properties of black lipid membranes.Z. Naturforsch. 31c:40
Papahadjopoulos, D., Poste, G. 1975. Calcium-induced phase separation and fusion in phospholipid membranes.Biophys. J. 15:945
Paul, K.G. 1947. Oxidation-reduction potential of cytochromec.Arch. Biochem. 12:441
Person, Ph., Zipper, H., Felton, J.H. 1969. Cytochrome oxidase solubilization at high pH.Arch. Biochem. Biophys. 131:457
Reich, M., Wainio, W.W. 1961a. A cytochromec-phospholipid complex.J. Biol. Chem. 236:3058
Reich, M., Wainio, W.W. 1961b. Role of phospholipids in cytochromec oxidase activity.J. Biol. Chem. 236:3062
Steinemann, A., Läuger, P. 1971. Interaction of cytochromec with phospholipid monolayers and bilayer membranes.J. Membrane Biol. 4:74
Takano, T., Kalli, O.B., Swanson, R., Dickerson, R.E. 1973. The structure of ferrocytochromec at 2.45 Å resolution.J. Biol. Chem. 248:5234
Theorell, H., Åkesson, Å. 1941. Studies on cytochromec. III. Titration curves.J. Am. Chem. Soc. 63:1820
White, S.H. 1970. A study of lipid bilayer membrane stability using precise measurements of specific capacitance.Biophys. J. 10:1127
Zöllner, N., Eberhagen, D. 1965. Untersuchung und Bestimmung der Lipoide im Blut. Springer-Verlag, Berlin
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Nöll, G.G. Simultaneous measurements of optical and electrical properties of artificial membranes composed of mitochondrial lipids and their interaction with cytochromec . J. Membrain Biol. 27, 335–346 (1976). https://doi.org/10.1007/BF01869144
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DOI: https://doi.org/10.1007/BF01869144