Binding of a tightly folded artificial mitochondrial precursor protein to the mitochondrial outer membrane involves a lipid-mediated conformational change.
An artificial mitochondrial precursor protein (the presequence of cytochrome oxidase subunit IV fused to mouse dihydrofolate reductase) binds to isolated yeast mitochondrial outer membranes and to liposomes whose phospholipid composition resembles that of outer membranes. In both cases, binding is strongly inhibited by low temperature or methotrexate (which stabilizes the dihydrofolate reductase moiety) and partly inhibited by adriamycin (which binds to acidic phospholipids). Binding is accompanied by partial unfolding of the protein. Binding of the urea-denatured fusion protein to outer membranes or liposomes is insensitive to low temperature, methotrexate, or adriamycin. These results, and those reported in the accompanying paper (Eilers, M., Endo, T., and Schatz, G. (1989) J. Biol. Chem. 264, 2945-2950) suggest that import of this fusion protein into isolated mitochondria involves at least partial unfolding by acidic phospholipids on the mitochondrial surface.
Endo
T
T
Biocenter, University of Basel, Switzerland.
Eilers
M
M
Schatz
G
G
D004317:Doxorubicin / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000378:metabolism*; D004792:Enzyme Precursors / Q000378:metabolism*; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D046911:Macromolecular Substances; D008563:Membrane Lipids / Q000502:physiology; D008928:Mitochondria / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism*
D004792:Enzyme Precursors; D008081:Liposomes; D046911:Macromolecular Substances; D008563:Membrane Lipids; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D004317:Doxorubicin; D013762:Tetrahydrofolate Dehydrogenase; D003576:Electron Transport Complex IV