Investigations on the insertion of the mitochondrial precursor protein apocytochrome c into model membranes.
Different aspects of the interaction of apocytochrome c and model membranes composed of negatively charged lipids, were studied in order to get insight into the nature of this interaction. The effect of the protein on the lipid packing properties are revealed by DSC, ESR and monolayer techniques. These experiments clearly demonstrate that upon electrostatic interaction with the negatively charged phospholipids, apocytochrome c is able to penetrate into the hydrophobic region of the model membrane. In the case of 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol, this results in a perturbation of 160 lipid molecules per apocytochrome c molecule. Most likely, apocytochrome c disrupts the formation of the gel phase and restricts the lipid chain motion above the gel to liquid-crystalline phase transition. Tryptophan fluorescence measurements confirm that at least a part of the protein penetrates into the bilayer, and suggest that after this penetration, the tryptophan (residue no. 59) is located in the glycerol backbone region of the phospholipids. Although the secondary structure of apocytochrome c is predicted to contain about 35% of alpha-helical structure, the CD pattern of an aqueous solution of the protein is featureless. However, negatively charged lipids are able to express this alpha-helical potency in the apocytochrome c, which might be important for the insertion of the protein into lipid membranes.
D000818:Animals; D001059:Apoproteins / Q000378:metabolism*; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D019826:Cholic Acid; D002793:Cholic Acids / Q000494:pharmacology; D002942:Circular Dichroism; D003574:Cytochrome c Group / Q000378:metabolism*; D045304:Cytochromes c; D004578:Electron Spin Resonance Spectroscopy; D005960:Glucosides / Q000494:pharmacology; D008433:Mathematics; D008563:Membrane Lipids / Q000378:metabolism*; D008567:Membranes, Artificial; D011312:Pressure; D011487:Protein Conformation; D013499:Surface Properties
D001059:Apoproteins; D002793:Cholic Acids; D003574:Cytochrome c Group; D005960:Glucosides; D008563:Membrane Lipids; D008567:Membranes, Artificial; C018619:octyl-beta-D-glucoside; D045304:Cytochromes c; D019826:Cholic Acid