Synthesis of Stably Charged Arg-tRNA Arg for Structural Analysis.

Posttranslational protein arginylation catalyzed by arginyl transferases is a mechanism to regulate multiple physiological processes. This protein arginylation reaction uses a charged Arg-tRNA Arg as the donor of arginine (Arg). The inherent instability of the ester linkage of the arginyl group to the tRNA, which is sensitive to hydrolysis at the physiological pH, makes it difficult to obtain structural information on how the arginyl transfer reaction is catalyzed. Here, we describe a methodology to synthesize stably charged Arg-tRNA Arg that would facilitate structural analysis. In the stably charged Arg-tRNA Arg , the ester linkage is replaced with an amide linkage, which is resistant to hydrolysis even at alkaline pH.