Efficient interaction of nonpolar lipids with intermediate filaments of the vimentin type.
Based on the finding that vimentin isolated and purified from cultured mammalian cells is heavily contaminated by neutral lipids, the binding of a series of radioactively labeled nonpolar lipids to pure, delipidated vimentin was investigated. Employing gel permeation chromatography of the complexes on Sephacryl S-300, cholesterol, cholesteryl fatty acid esters and mono-, di- and triglycerides were found to efficiently associate with vimentin. These compounds also showed a strong tendency to bind to vimentin filaments. While the non-alpha-helical head piece of vimentin did not interact with neutral lipids under the above assay conditions, the alpha-helical rod domain was highly active. When cholesterol or 1,2-dioleoyl-glycerol was incorporated into phospholipid vesicles, the affinity of the liposomes for vimentin filaments was considerably increased. However, in sucrose density gradient equilibrium centrifugation the filament-vesicle adducts were only stable when the liposomes contained negatively charged phospholipids. These results suggest that the association of intermediate filaments with lipid vesicles is initiated by interaction of the arginine-rich N-termini of their subunit proteins with the negatively charged vesicle surface and stabilized by partial insertion of the protein molecules into the lipid bilayer, particularly at those sites where immiscible, nonpolar lipids create defects in phospholipid packing. Very likely, nonpolar lipids play a significant role in the interaction of intermediate filaments with natural membrane systems.
D000818:Animals; D002250:Carbon Radioisotopes; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D007382:Intermediate Filaments / Q000378:metabolism* / Q000648:ultrastructure; D050356:Lipid Metabolism; D051379:Mice; D008854:Microscopy, Electron; D014316:Tritium; D014746:Vimentin / Q000302:isolation & purification / Q000378:metabolism*
D002250:Carbon Radioisotopes; D005227:Fatty Acids; D005230:Fatty Acids, Nonesterified; D014746:Vimentin; D014316:Tritium; D002784:Cholesterol