Targeting phosphodiesterases as a strategy for killing tumor cells

Abstract

ribonucleases (RNases) are being employed as alternative cytotoxic proteins to the conventionally used ones such as ricin andPseudomonas exotoxin. Mammalian RNases are attractive enzymes because of their comparable cytotoxicity when suitably directed and the likelihood of lower immunogenicity compared to plant and bacterial toxins. Bovine seminal RNase (BSRNase) is a member of the RNase superfamily, but differs in many interesting ways. Unlike the rest of the family it is dimeric, and possesses antitumor and immunosupressive properties. These features make it a choice candidate for a single-chain antibody (scFv) based immunotoxin. This work describes preliminary data on the construction, expression inEscherichia coli and characterization of a tumor-specific scFv (directed against human placental alkaline phosphatase)-BSRNase chimeric molecule. It is shown that the created molecule has RNA degrading activity and antigen-binding activity when refolded from bacterial inclusion bodies.