Protein phosphatase 2A is associated in an inactive state with microtubules through 2A1-specific interaction with tubulin.

Protein phosphatase (PP) 2A1, a trimer composed of A-, B- and C-subunits in the PP2A family, has been regarded as a principal form localizing at microtubules (MT), but PP2A2, the dimer of A- and C-subunits, has not. Substantiating the claim, the present work shows that the PP2A1 but not PP2A2, both isolated from bovine extract, largely associated with the purified preparation of MT. Furthermore, PP2A1 was found to bind purifiedtubulin polymerized by taxol. The presence of MT associated proteins with purified tubulin hardly affected the binding of PP2A1 to the tubulin. In addition, PP2A1 activity towards glycogen phosphorylase, a probably unphysiological but good substrate, was similarly inhibited by MT proteins and purified tubulin, which accounts for > or =85% of MT proteins, with their IC(50) of about 0.15 mg/ml. In contrast, the inhibition of PP2A2 was about 40% with 1 mg/ml MT proteins and 20% with 0.8 mg/ml tubulin, consistent with its weak association with MT. Therefore, the association with and resultant inhibition by MT proteins of PP2A1 is largely effected by the binding of PP2A1 to tubulin molecule. Moreover, PP2A1 isolated from MT has higher affinity for polymerized MT proteins than has PP2A1 from the postmicrotubule supernatant. The MT PP2A1 has also higher sensitivity to the inhibition by tubulin and MT proteins than has the supernatant PP2A1 (IC(50): 0.1-0.2 mg/ml vs. 0.3-0.6 mg/ml), demonstrating the importance of its association with polymerized tubulin.