Phosphorylation in vitro of vimentin by protein kinases A and C is restricted to the head domain. Identification of the phosphoserine sites and their influence on filament formation.
The in vitro phosphorylation of vimentin, the intermediate filament protein of mesenchymal cells, by kinases A and C is serine-specific and involves only the N-terminal head domain. In oligomeric protofilament units each kinase recognizes five sites, which have been identified by sequence analysis. Kinase C introduces 1.5 mol phosphate/mol vimentin, while kinase A treatment results in 4 mol phosphate/mol. Kinase-A-treated oligomers do not polymerize in standard assays whereas kinase C treatment has no inhibitory effect. Filaments exposed to kinase A remain stable and incorporate only 1.7 mol phosphate/mol vimentin. These phosphates are essentially restricted to two of the five kinase A sites found in protofilament units. Thus the head domain, previously related to in vitro assembly competence and filament stability, changes in accessibility between the oligomeric and polymeric state. We discuss the possibility that in vivo phosphorylation of vimentin filaments by kinase A may not necessarily be accompanied by an extensive depolymerization. It could instead involve a dynamic change of the filament surfaces, which could alter the interaction of the filaments with other cellular structures.
Geisler
N
N
Max Planck Institute for Biophysical Chemistry, Göttingen, Federal Republic of Germany.
Hatzfeld
M
M
Weber
K
K
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D012694:Serine / Q000031:analogs & derivatives*; D013552:Swine; D013917:Thrombin / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism*
D046911:Macromolecular Substances; D010446:Peptide Fragments; D010710:Phosphates; D014746:Vimentin; D010768:Phosphoserine; D012694:Serine; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D011493:Protein Kinase C; D013917:Thrombin