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Comparison of the phosphorylation of microtubule-associated protein tau by non-proline dependent protein kinases

Abstract

Microtubule-associated protein tau from Alzheimer brain has been shown to be phosphorylated at several ser/thr-pro and ser/thr-X sites (Hasegawa, M. et al., J. Biol. Chem, 267, 17047–17054, 1992). Several proline-dependent protein kinases (PDPKs) (MAP kinase, cdc2 kinase, glycogen synthase kinase-3, tubulin-activated protein kinase, and 40 kDa neurofilament kinase) are implicated in the phosphorylation of the ser-thr-pro sites. The identity of the kinase(s) that phosphorylate that ser/thr-X sites are unknown. To identify the latter kinase(s) we have compared the phosphorylation of bovine tau by several brain protein kinases. Stoichiometric phosphorylation of tau was achieved by casein kinase-1, calmodulin-dependent protein kinase II, Gr kinase, protein kinase C and cyclic AMP-dependent protein kinase, but not with casein kinase-2 or phosphorylase kinase. Casein kinase-1 and calmodulin-dependent protein kinase II were the best tau kinases, with greater than 4 mol and 3 mol32P incorporated, respectively, into each mol of tau. With the sequential addition of these two kinases,32P incorporation approached 6 mol. Peptide mapping revealed that the different kinases largely phosphorylate different sites on tau. After phosphorylation by casein kinase-1, calmodulin-dependent protein kinase II, Gr kinase, cyclic AMP-dependent protein kinase and casein kinase-2, the mobility of tau isoforms as detected by SDS-PAGE was decreased. Protein kinase C phosphorylation did not produce such a mobility shift. Our results suggest that one or more of the kinases studied here may participate in the hyperphosphorylation of tau in Alzheimer disease. Such phosphorylation may serve to modulate the activaties of other tau kinases such as the PDPKs.

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Abbreviations

PHF:

paired helical filaments

A-kinase:

cyclic AMP-dependent protein kinase

CaM kinase II:

calcium/calmodulin-dependent protein kinase II

C-kinase:

calcium-phospholipid-dependent protein kinase

CK-1:

casein kinase-1

CK-2:

casein kinase-2

Gr kinase:

calcium/calmodulin-dependent protein kinase from rat cerebellum

GSK-3:

glycogen synthase kinase-3

MAP kinase:

mitogen-activated protein kinase

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Correspondence to Khalid Iqbal.

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Singh, J., Grundke-Iqbal, I., McDonald, B. et al. Comparison of the phosphorylation of microtubule-associated protein tau by non-proline dependent protein kinases. Mol Cell Biochem 131, 181–189 (1994). https://doi.org/10.1007/BF00925955

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Key words

  • tau phosphorylation
  • microtubules
  • protein kinases
  • Alzheimer disease
  • Tau-1 antibody