The interactions of bovine brain MAP2 and tau, recombinant murine MAP2C, and recombinant human tau with phosphatidylinositol vesicles yield apparent Kd values of 51 +/- 6 nM, 2.4 +/- 0.6 microM, 1.4 +/- 0.1 microM, and 1.6 +/- 0.2 microM, respectively. Examinations of the binding of MAP2 and/or MAP2C to phosphatidylcholine vesicles doped with phosphatidylinositol or to phosphatidylserine vesicles and of thrombin-digested MAP2C to phosphatidylinositol vesicles demonstrates that the observed high affinity of the MAP2: phosphatidylinositol binding is due to the contributions of two separate interactions. A low-affinity site (Kd = 1.5-2.5 microM) is located within the C-terminal domain and affects the nonspecific interaction of MAP2, MAP2C, and tau with anionic phospholipids. The second site, with an apparent Kd of 221 +/- 25 nM, is located within the MAP2-specific peptide, which is eliminated from MAP2C by differential gene splicing. It is proposed that the high affinity of MAP2 for phosphatidylinositol contributes to the spatial and temporal regulation of the dendritic cytoskeleton.