Proteolytic enzymes seem to play important roles in the life cycles of all medically important protozoan parasites, including the organisms that cause malaria, trypanosomiasis, leishmaniasis, amebiasis, toxoplasmosis, giardiasis, cryptosporidiosis and trichomoniasis. Proteases from all four major proteolytic classes are utilized by protozoans for diverse functions, including the invasion of host cells and tissues, the degradation of mediators of the immune response and the hydrolysis of host proteins for nutritional purposes. The biochemical and molecular characterization of protozoan proteases is providing tools to improve our understanding of the functions of these enzymes. In addition, studies in multiple systems suggest that inhibitors of protozoan proteases have potent antiparasitic effects. This review will discuss recent advances in the identification and characterization of protozoan proteases, in the determination of the function of these enzymes, and in the evaluation of protease inhibitors as potential antiprotozoan drugs.