myo-Inositol monophosphatase (E.C. 3.1.3.25) hydrolyzes inositol monophosphate to form free myo-inositol, the precursor for the inositol phospholipid second-messenger signaling systems. The biochemical properties of the enzyme were examined in detail. 2. The enzyme exhibited significant hydrolytic activity only on phosphotyrosine among physiological substrates tested in the presence of Zn2+ ions in an acidic environment. 3. The enzyme was recognized and immunoprecipitated with polyclonal antibodies developed against the Zn2+-dependent tyrosine phosphatase of bovine brain. 4. These results indicate that myo-inositol monophosphatase exhibits Zn2+-dependent tyrosine phosphatase activity in an acidic environment and has immunological identity with a Zn2+-dependent tyrosine phosphatase.