Studying Reversible Protein Post‐translational Modification through Co‐translational Modification

Abstract

Understanding the post-translational modifications of targeted proteins is of great significance for manipulating the physiological processes of eukaryotes. Chemical biology tools have been used to investigate the biological roles of those posttranslational modifications at particular sites, especially genetic code expansion technology, which can also be combined with the concept of synthetic biology to generate a genetically modified organism with a synthetic auxotroph for co-translational modification components. In this concept, we will introduce applications, limitations, and perspectives of genetic code expansion technology for studying post-translational modification based on recent progresses. Future perspectives of genetically modified organisms also will be discussed in regard to the application of post-translational modification research.

Orthogonal Translation for Site-Specific Installation of Post-translational Modifications

“…To fill in this gap, the genetic code expansion (GCE) strategy has been applied for PTM studies. − It introduces an orthogonal translation system (OTS) into living cells to genetically encode a noncanonical amino acid (ncAA) at a chosen site of target proteins. − The OTS is mainly composed of an engineered aminoacyl-tRNA synthetase (AARS), which recognizes the ncAA, and an engineered tRNA, which bears a specific anticodon to read an assigned codon in mRNA (Figure ). − The orthogonality of an OTS includes the following: First, the target ncAA cannot be recognized by host AARSs; Second, the engineered AARS cannot recognize canonical amino acids and cannot aminoacylate host tRNAs; Third, the engineered tRNA cannot be aminoacylated by host AARSs.…”