Correspondence to E. Mandelkow, Max-Planck-Unit for Structural Molecular Biology, Notkestr. 85, D-22607 Hamburg, Germany
Sequential phosphorylation of Tau by glycogen synthase kinase-3β and protein kinase A at Thr212 and Ser214 generates the Alzheimer-specific epitope of antibody AT100 and requires a paired-helical-filament-like conformation
Article first published online: 25 DEC 2001
European Journal of Biochemistry
Volume 252, Issue 3, pages 542–552, March (II) 1998
How to Cite
Zheng-Fischhöfer, Q., Biernat, J., Mandelkow, E.-M., Illenberger, S., Godemann, R. and Mandelkow, E. (1998), Sequential phosphorylation of Tau by glycogen synthase kinase-3β and protein kinase A at Thr212 and Ser214 generates the Alzheimer-specific epitope of antibody AT100 and requires a paired-helical-filament-like conformation. European Journal of Biochemistry, 252: 542–552. doi: 10.1046/j.1432-1327.1998.2520542.x
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Abbreviations. GSK, glycogen synthase kinase; PKA, protein kinase A; PHF, paired helical filament; ISPK, insulin-stimulated protein kinase; MAP kinase, mitogen-activated protein kinase; PKC, protein kinase C; Cdk, cyclin-dependent kinase.
- Issue published online: 25 DEC 2001
- Article first published online: 25 DEC 2001
- (Received 30 September/11 December 1997)
- Cited By
- microtubule-associated protein;
- epitope mapping.
AT100 is a monoclonal antibody highly specific for phosphorylated Tau in Alzheimer paired helical filaments. Here we show that the epitope is generated by a complex sequence of sequential phosphorylation, first of Ser199, Ser202 and Thr205 (around the epitope of antibody AT8), next of Thr212 by glycogen synthase kinase (GSK)-3β (a proline-directed kinase), then of Ser214 by protein kinase A (PKA). Conversely, if Ser214 is phosphorylated first it protects Thr212 and the Ser-Pro motifs around the AT8 site against phosphorylation, and the AT100 epitope is not formed. The generation of the AT100 epitope requires a conformation of tau induced by polyanions such as heparin, RNA or poly(Glu), conditions which also favor the formation of paired helical filaments. The Alzheimer-like phosphorylation can be induced by brain extracts. In the extract, the kinases responsible for generating the AT100 epitope are GSK-3β and PKA, which can be inhibited by their specific inhibitors LiCl and RII, respectively. A cellular model displaying the reaction with AT100 is presented by Sf9 insect cells transfected with Tau. Knowledge of the events and kinases generating the AT100 epitope in cells might allow us to study the degeneration of the cytoskeleton in Alzheimer's disease.