Influence of surface charge on the incorporation and orientation of cytochrome c oxidase in liposomes

AbstractCytochrome c-oxidase is usually oriented 80–90% right-side-out when reconstituted with asolectin by the cholate dialysis method. Transformation of positively charged lysine groups at the matrix domain into negatively charged groups with succinic anhydride results in random orientation. A random orientation is also found after reconstitution in phosphatidylcholine, which can be changed into predominant right-side-out orientation by addition of cardiolipin. It is concluded that electrostatic interaction between positively charged groups of cytochrome c-oxidase with negative groups of phospholipids determines the asymmetric orientation of the enzyme in liposomes.Liposome; Cytochrome c-oxidase; Orientation; Protein modification; Succinic anhydride; Cardiolipi