Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins

Irem Avcilar-Kucukgoze; Howard Gamper; Ya-Ming Hou; Anna Kashina

STAR Protocols (Dec 2020)

Purification and Use of tRNA for Enzymatic Post-translational Addition of Amino Acids to Proteins

Irem Avcilar-Kucukgoze,
Howard Gamper,
Ya-Ming Hou,
Anna Kashina

Affiliations

Irem Avcilar-Kucukgoze: Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19144, USA; Corresponding author
Howard Gamper: Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19144, USA
Ya-Ming Hou: Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19144, USA
Anna Kashina: Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Corresponding author

Journal volume & issue: Vol. 1, no. 3
p. 100207

Abstract

Read online

Summary: Post-translational addition of amino acids to proteins by enzymes using aminoacyl-tRNA is an emerging regulatory mechanism. Examples include Arg transfer in eukaryotes, Leu/Phe transfer in bacteria, and tRNA-synthetase-mediated addition of amino acids to Lys side chains. Here, we present a method of purification and use of tRNA for such reactions, focusing on tRNAArg and its use for arginylation. This method can also be used for other tRNA-mediated reactions.For complete details on the use and execution of this protocol, please refer to Avcilar-Kucukgoze et al. (2020)

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords