Assaying ATE1 Activity In Vitro

Junling Wang; Anna S Kashina

doi:10.1007/978-1-0716-2942-0_15

Assaying ATE1 Activity In Vitro

Methods Mol Biol. 2023:2620:113-117. doi: 10.1007/978-1-0716-2942-0_15.

Authors

Junling Wang¹, Anna S Kashina²

Affiliations

¹ University of Pennsylvania, Philadelphia, PA, USA.
² Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA. akashina@upenn.edu.

PMID: 37010756
DOI: 10.1007/978-1-0716-2942-0_15

Abstract

Here, we describe a standard arginyltransferase assay in vitro using bacterially expressed purified ATE1 in a system with a minimal number of components (Arg, tRNA, Arg-tRNA synthetase, and arginylation substrate). Assays of this type have first been developed in the 1980s using crude ATE1 preparations from cells and tissues and then perfected recently for the use with bacterially expressed recombinant protein. This assay represents a simple and efficient way to measure ATE1 activity.

Keywords: ATE1; Arginylation; Arginylation assay.

MeSH terms

Aminoacyltransferases* / genetics
Arginine / metabolism
Protein Processing, Post-Translational*
Recombinant Proteins / genetics
Recombinant Proteins / metabolism

Substances

Aminoacyltransferases
Recombinant Proteins
Arginine