Synthesis of Stably Charged Arg-tRNAArg for Structural Analysis

Abstract

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.

Keywords: Arg-tRNAArg; Arginylation; Structural Analysis; tRNA.