Alzheimer disease specific phosphoepitopes of Tau interfere with assembly of tubulin but not binding to microtubules.
In Alzheimer disease (AD)-affected neurons, the Tau protein is found in an aggregated and hyperphosphorylated state. A common hypothesis is that Tau hyperphosphorylation causes its dissociation from the microtubular surface, with consequently a breakdown of the microtubules (MTs) and aggregation of the unbound Tau. We evaluated the effect of Tau phosphorylation on both tubulin assembly and MT binding. We show that the cyclin-dependent kinase 2/cyclin A3 kinase complex can generate the AT8 and AT180 AD-specific phospho-epitopes and use NMR spectroscopy to validate qualitatively and quantitatively the phospho content of our samples. The simultaneous presence of both epitopes disables the tubulin assembly capacity of Tau in conditions whereby Tau is the driving force for the assembly process but does not, however, inhibit MT assembly when the latter is driven by an increased tubulin concentration. When compared to the isolated MT binding repeats (K(d)=0.3 microM), the phospho-Tau retains a substantial affinity for preformed MTs (K(d)=11 nM), suggesting that the phosphorylated proline-rich region still participates in the binding event. Our results hence indicate that the sole phosphorylation at the AT8/AT180 epitopes, although leading to a functional defect for Tau, is not sufficient for its dissociation from the MT surface and subsequent aggregation as observed in AD.
Amniai
Laziza
L
Université des Sciences et Technologies de Lille 1, Villeneuve d'Ascq, France.
Barbier
Pascale
P
Sillen
Alain
A
Wieruszeski
Jean-Michel
JM
Peyrot
Vincent
V
Lippens
Guy
G
Landrieu
Isabelle
I
D000544:Alzheimer Disease / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D000939:Epitopes; D006801:Humans; D008870:Microtubules / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation; D011485:Protein Binding / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D014404:Tubulin / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism*
D000939:Epitopes; D011994:Recombinant Proteins; D014404:Tubulin; D016875:tau Proteins; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2