Constructing Photoactivatable Protein with Genetically Encoded Photocaged Glutamic Acid

Abstract

Genetically replacing an essential residue with the corresponding photocaged analogues via genetic code expansion (GCE) constitutes a useful and unique strategy to directly and effectively generate photoactivatable proteins. However, the application of this strategy is severely hampered by the limited number of encoded photocaged proteinogenic amino acids. Herein, we report the genetic incorporation of photocaged glutamic acid analogues in E. coli and mammalian cells and demonstrate their use in constructing photoactivatable variants of various fluorescent proteins and SpyCatcher. We believe genetically encoded photocaged Glu would significantly promote the design and application of photoactivatable proteins in many areas.

A DNA Force Circuit for Exploring Protein‐Protein Interactions at the Single‐Molecule Level†

“…Genetic code expansion technology, on the other hand, allows for the introduction of unnatural amino acids into proteins, providing enhanced biocompatibility and biological orthogonality while enabling site‐specific and stoichiometric control over conjugation reactions. [ 29 ] Through atomic force microscopy morphology analysis, we were able to evaluate the critical steps in the development of methods during the preparation and assembly of PPI structures. After optimizing the preparation and assembly procedure of single‐molecule PPI structures in this study, the efficiency of bioconjugate reactions has been improved, as well as the efficiency of protein engineering using expanded genetic codes to introduce unnatural amino acids.…”

SpyTag Peptide with Alkoxyl Aspartic Acids for pH-Dependent Activation of the SpyCatcher/Tag System

“…In this system, a light-oxygen-voltage sensing domain, the so-called LOV domain, which converts the structure upon blue light exposure, was fused to SpyTag to enable blue light-dependent activation. In addition, there are several reports for the photoactivation systems using photocaged amino acids introduced in SpyCatcher to enable light-induced activation. − By using such a photoactivation system, the spatiotemporal complex formation in a cell is possible.…”