[Association of microtubule promoting and binding activity of tau with its phosphorylation sites].
OBJECTIVE
To explore the association between the abnormal phosphorylation sites found in Alzheimer disease (AD) tau and the inhibition of its biological activity.


METHODS
Ultracentrifugation, chromatography, manual Edman degradation and autosequence techniques were used to prepare and phosphorylate human recombinant tau, isolate and purify 32P tau peptides and determine phosphorylation sites.


RESULTS
(1) Phosphorylation of tau by casein kinase-1 (CK-1), cyclic AMP-dependent protein kinase (PKA) and glycogen synthetase kinase-3 (GSK-3) differentially inhibited its biological activity, and the inhibition of this activity of tau by GSK-3 was significantly increased if tau was prephosphorylated by CK-1 or PKA. The most potent inhibition was seen by a combined phosphorylation of tau with PKA and GSK-3. (2) The treatment of tau by PKA and GSK-3 combination induced phosphorylation of tau at Ser-195, Ser-198, Ser-199, Ser-202, Thr-205, Thr-231, Ser-235, Ser-262, Ser-356, Ser-404, whereas only Thr-181, Ser-184, Ser-262, Ser-356 and Ser-400 were phosphorylated by GSK-3 alone under the same conditions. Among the above-mentioned phosphorylation sites, Ser-198, Ser-199, Ser-202, Thr-205, Thr-231, Ser-235, Ser-262, Ser-400 and Ser-404 were seen in Alzheimer tau. The phosphorylation of Ser-262 only slightly inhibited its biological activity, and Ser-198, Ser-199, Ser-202, Thr-231, Ser-235, Ser-400 and Ser-404 also presented in fetal tau which was highly active, suggesting that Thr-205 was the unique site which both caused the potent inhibition of biological activity and specifically presented in AD abnormally phosphorylated tau.


CONCLUSIONS
Phosphorylation of Thr-205 might play a key role in tau pathology in AD.
Wang
J
J
Department of Pathophysiology, Tongji Medical University, Wuhan 430030, China. bswzl@tjmu.edu.cn
Wei
Z
Z
Wang
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Q
Grundke-Iqbal
I
I
Iqbal
K
K
D000544:Alzheimer Disease / Q000201:enzymology / Q000378:metabolism*; D001665:Binding Sites; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008870:Microtubules / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism* / Q000502:physiology
D011994:Recombinant Proteins; D016875:tau Proteins; D011505:Protein-Tyrosine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D038362:Glycogen Synthase Kinase 3