Incorporation of mitochondrial membrane proteins into liposomes containing acidic phospholipids

• Published in: The Journal of biological chemistry Vol. 251; no. 21; pp. 6831 - 6837
• Main Authors: Eytan, G D, Matheson, M J, Racker, E
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 10.11.1976
• Subjects: Adenosine Triphosphatases - metabolism; Calcium - pharmacology; Darkness; Edetic Acid - pharmacology; Electron Transport Complex IV - metabolism; Enzyme Activation - drug effects; Halobacterium - metabolism; Kinetics; Light; Liposomes; Magnesium - pharmacology; Membrane Proteins - metabolism; Mitochondria - metabolism; Oligomycins - pharmacology; Phosphatidylinositols - pharmacology; Phosphatidylserines - pharmacology; Phospholipids - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)33019-3

Cytochrome oxidase, QH2-cytochrome c reductase, and the oligomycin-sensitive adenosine triphosphatase were incorporated into liposomes by a new procedure which yielded unidirectional orientation of the proteins. Cytochrome oxidase was reconstituted in the mitochrondrial orientation and the adenosine triphosphatase in the submitochondrial orientation. Reconstitutions were achieved by incubating the proteins at room temperature with liposomes which contained phosphatidylcholine, phosphatidylethanolamine, and an acidic phospholipid (cardiolipin, phosphatidylinositol, or phosphatidylserine). The incorporation occurred without added detergent or sonication. This incorporation procedure may serve as a model for the insertion of proteins in vivo.