The intermediate-filament proteins vimentin and desmin are phosphorylated in specific domains.
Analysis of specific fragments of vimentin and desmin from 32P-labeled BHK-21 cells indicated that these intermediate-filament subunit proteins are phosphorylated in specific regions or domains. High performance liquid chromatography and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis of lysine-specific protease-generated fragments demonstrated that both molecules were phosphorylated in their amino terminal or "head" domains. While this was the predominant site of phosphorylation for vimentin, additional phosphorylated fragments from desmin were observed. Chemical cleavage of [32P]desmin and subsequent examination of the phosphorylated peptides indicated that the major site of desmin phosphorylation was located within the "tail" domain. Analysis of vimentin and desmin from non-mitotic and mitotically selected cells indicated that the increased phosphorylation of intermediate-filament proteins observed during cell division occurs within the amino terminal domains of both molecules. These studies indicate that the increased phosphorylation of filament proteins during mitosis may involve the function of the amino terminal domain. In addition, filament proteins may be phosphorylated in a subunit-protein-specific manner which may reflect subunit-specific functions.
Evans
R M
RM
Department of Pathology, University of Colorado Health Sciences Center, Denver 80262.
D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003599:Cytoskeleton / Q000648:ultrastructure; D003893:Desmin / Q000302:isolation & purification; D007739:L Cells / Q000166:cytology; D051379:Mice; D010447:Peptide Hydrolases; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000302:isolation & purification; D010766:Phosphorylation; D014746:Vimentin / Q000302:isolation & purification
D003893:Desmin; D010748:Phosphopeptides; D014746:Vimentin; D010447:Peptide Hydrolases