Synthesis of Peptides and Proteins with Site-Specific Glutamate Arginylation

Abstract

Solid-phase peptide synthesis and protein semi-synthesis are powerful methods for site-specific modification of peptides and proteins. We describe protocols using these techniques for the syntheses of peptides and proteins bearing glutamate arginylation (E^Arg) at specific sites. These methods overcome challenges posed by enzymatic arginylation methods and allow for a comprehensive study of the effects of E^Arg on protein folding and interactions. Potential applications include biophysical analyses, cell-based microscopic studies, and profiling of E^Arg levels and interactomes in human tissue samples.

Keywords: Glutamate arginylation; Native chemical ligation; Posttranslational modification; Protein semi-synthesis; Solid-phase peptide synthesis.