Global analysis of post-translational sidechain arginylation using pan-arginylation antibodies

Mol Cell Proteomics. 2023 Oct 11:100664. doi: 10.1016/j.mcpro.2023.100664. Online ahead of print.ABSTRACTArginylation is a post-translational modification mediated by the arginyltransferase ATE1, which transfers the amino acid arginine to a protein or peptide substrate from a tRNA molecule. Initially, arginylation was thought to occur only on N-terminally exposed acidic residues, and its function was thought to be limited to targeting proteins for degradation. However, more recent data has shown that ATE1 can arginylate sidechains of internal acidic residues in a protein without necessarily affecting metabolic stability. This greatly expands the potential targets and functions of arginylation, but tools for studying this process have remained limited. Here, we report the first global screen specifically for sidechain arginylation. We generate and validate "pan-arginylation" antibodies, which are designed to detect sidechain arginylation in any amino acid sequence context. We use these antibodies for immunoaffinity enrichment of sidechain arginylated proteins from wildtype and Ate1 knockout cell lysates. In this way, we identify a limited set of proteins that likely undergo ATE1-dependent sidechain arginylation and that are enriched in specific cellular roles, including translation, splicing, and the cytoskeleton.PMID:37832787 | DOI:10.1016/j.mcpro.2023.100664
Source: Molecular and Cellular Proteomics : MCP - Category: Molecular Biology Authors: Source Type: research