Methods to Study Arginylation of Tubulin

Arginylation is a posttranslational modification in which arginine is transferred from a tRNA molecule onto a protein by arginyltransferase 1 (ATE1). Previous studies have found that Ate1 knockout causes cytoskeletal defects at both the cellular and organismal levels, but the protein targets of arginylation that mediate these effects remain poorly defined. Some of the Ate1 knockout phenotypes have been linked to β-actin arginylation, but more recently, the role of tubulin arginylation has become an active area of study. Here, we describe methods for studying the arginylation of tubulin both in vitro and from cell/tissue extracts.