We previously showed that exposure to metal-laden combustion particles disregulates protein tyrosine phosphate homeostasis in human airway epithelial cells (HAEC). More recently, we reported that exposure to certain metal ions activates mitogen-activated protein kinases in HAEC. To study the mechanism responsible, we examined the effects of arsenic (As), vanadium (V), and zinc (Zn) on tyrosine phosphate catabolism in BEAS S6 cells or cultured human bronchial epithelial cells. Western blots and immunocytochemical analyses showed that exposure to noncytotoxic levels of As, V, or Zn resulted in increased levels of protein phosphotyrosines in HAEC. Tyrosine phosphatase activity, measured against [(32)P]-labeled PolyGlu:Tyr, was markedly inhibited in cells treated with V or Zn but was unaffected by exposure to As. Fast performance liquid chromatography fractionation and subsequent in-gel phosphatase activity assay of HAEC protein extracts revealed the presence of numerous tyrosine phosphatases, of varying molecular weights, that were effectively inhibited by exposure to V or Zn ions. As had no discernible effect on these enzymes. The protein tyrosine phosphatase PTP1B, immunoprecipitated from HAEC, was similarly inhibited by V and Zn but not by As ions. These data show that V and Zn may induce tyrosine phosphate accumulation by inhibiting dephosphorylation and implicate kinase activation as the mechanism in HAEC exposed to As. These findings suggest that metal exposure can activate signaling pathways through multiple mechanisms.