Heteropolymer filaments of vimentin and desmin in vascular smooth muscle tissue and cultured baby hamster kidney cells demonstrated by chemical crosslinking.
Certain smooth muscle cells of blood vessel walls as well as cultured baby hamster kidney cells contain simultaneously two different intermediate-sized filament (IF) proteins, desmin and vimentin. We have examined the question of the occurrence of both proteins in the same IF by chemically crosslinking the single cysteine group present in each of them. Oxidative crosslinking of filaments present in cytoskeletal preparations with cupric ion complexes of 1,10-phenanthroline resulted in formation of three types of dimers: vimentin-vimentin, desmin-desmin, and vimentin-desmin. These dimers were separated by NaDodSO4/polyacrylamide gel electrophoresis and characterized by binding of specific antibodies, by one- and two-dimensional gel electrophoresis of monomers obtained after cleavage of the disulfide bond by thiol agents, and by mapping of radioiodinated tryptic peptides. The demonstration of heterodimers of vimentin and desmin in vascular smooth muscle tissue of cow and chicken and in baby hamster kidney cells shows that the two proteins can be integrated in the same IF and can be nearest neighbors, oriented with their cysteine residues in a mirror-image symmetry. The possible existence of heteropolymer IF in other cell types is discussed.
D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D003432:Cross-Linking Reagents; D003599:Cytoskeleton / Q000648:ultrastructure*; D003893:Desmin; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000648:ultrastructure; D009124:Muscle Proteins / Q000378:metabolism*; D009130:Muscle, Smooth / Q000648:ultrastructure*; D014746:Vimentin
D003432:Cross-Linking Reagents; D003893:Desmin; D009124:Muscle Proteins; D014746:Vimentin