Influence of the phosphorylation state of neurofilament proteins on the interactions between purified filaments in vitro.
The extensive enzymic dephosphorylation of neurofilaments determined the progressive loss of their capacity to interconnect in vitro into a reticulated network, measured by the formation of highly viscous gels in purified preparations of neurofilaments [Leterrier & Eyer (1987) Biochem. J. 245, 93-101]. Conversely, a cyclic AMP-dependent activation of the gelation process was obtained by phosphorylation of the neurofilament proteins by the cyclic-nucleotide-dependent protein kinase added to the preparation. These findings argue for a direct relationship between the high phosphorylation level of the neurofilament subunits and the cross-bridging of the polymers in vitro. However, a transient stimulation of the neurofilament viscosity kinetics was also observed during the early steps of dephosphorylation with acid phosphatase, which, moreover, disappeared with longer incubation times before the net inhibition was obtained. In the same way, the calmodulin-dependent brain phosphatase, calcineurin, induced a permanent activation of the phenomenon, correlated with a low dephosphorylation capacity of the neurofilament molecules. Taken together, these results suggest a functional heterogeneity of the numerous phosphate groups of the neurofilament subunits and raise the hypothesis of a highly controlled regulation of the neurofilament cross-bridging by selective phosphorylation-dephosphorylation mechanisms.
Eyer
J
J
Centre de Neurochimie du C.N.R.S., Strasbourg, France.
Leterrier
J F
JF
D000135:Acid Phosphatase / Q000494:pharmacology; D000818:Animals; D019703:Calcineurin; D002147:Calmodulin / Q000494:pharmacology; D002148:Calmodulin-Binding Proteins / Q000494:pharmacology; D002417:Cattle; D000242:Cyclic AMP / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D007382:Intermediate Filaments / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D010749:Phosphoprotein Phosphatases / Q000494:pharmacology; D010766:Phosphorylation; D011494:Protein Kinases / Q000494:pharmacology; D014783:Viscosity
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D007381:Intermediate Filament Proteins; D000242:Cyclic AMP; D011494:Protein Kinases; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D000135:Acid Phosphatase