Orientation of the alpha-helices of apocytochrome c and derived fragments at membrane interfaces, as studied by circular dichroism.
The orientation of the different helical regions of the mitochondrial precursor protein apocytochrome c has been studied using circular dichroism on isolated fragments of this protein associated with oriented films composed of various phospholipids [de Jongh, H. H. J., Goormaghtigh, E., & Killian, J. A. (1994) Biochemistry (preceding article in this issue)]. Both the N and C terminus adopt helical structures in a membrane environment. The middle region can also be helical, but only in the presence of the N-terminal domain of the protein. In the presence of the unsaturated lipids dioleoylphosphatidylcholine and dioleoylphosphatidylglycerol, all three helices are found to have a preferred orientation perpendicular to the membrane normal, whereas in the presence of the saturated lipids dimyristoylphosphatidylcholine and dimyristoylphosphatidylglycerol, the terminal helices are preferentially oriented parallel to the membrane normal. In films composed of dioleoylphosphatidylserine, it is found that the N-terminal helix is oriented preferentially perpendicular, whereas the C-terminal helix is aligned more parallel to the membrane normal. The differences in preferred orientation between the terminal helices are demonstrated by molecular modeling of the helices at a water-lipid interface. The results are discussed in light of the translocation of apocytochrome c over the outer mitochondrial membrane, an important step in the import process of this protein in mitochondria.
de Jongh
H H
HH
Department of Biochemistry of Membranes, University of Utrecht, The Netherlands.
Brasseur
R
R
Killian
J A
JA
D000818:Animals; D001059:Apoproteins / Q000648:ultrastructure*; D002942:Circular Dichroism; D003198:Computer Simulation; D003574:Cytochrome c Group / Q000648:ultrastructure*; D045304:Cytochromes c; D006736:Horses; D066298:In Vitro Techniques; D008563:Membrane Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000648:ultrastructure*; D008567:Membranes, Artificial; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry; D017433:Protein Structure, Secondary; D014867:Water / Q000737:chemistry
D001059:Apoproteins; D003574:Cytochrome c Group; D008563:Membrane Lipids; D008565:Membrane Proteins; D008567:Membranes, Artificial; D010446:Peptide Fragments; D014867:Water; D045304:Cytochromes c