RNA molecules that bind to and inhibit the active site of a tyrosine phosphatase

Abstract

Protein tyrosine phosphatases (PTPases) are essential proteins in many cellular processes. In vitro selection was used to evolve high affinity RNA aptamers to the Yersinia PTPase from two random pools varying in length. Selected aptamers from the two different pools share a 21-residue conserved sequence. They bind to their target with dissociation constants of 18 and 28 nM and inhibit the enzyme with IC50 values of 10 and 35 nM, but do not bind a related PTPase. Modification of the PTPase's active site cysteine with the alkylating agent iodoacetate results in a loss of binding affinity. These experiments suggest that the selected aptamers act by binding at or near the active site and might therefore be useful in defining the interactions between PTPases and their targets.