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.

Keywords: Genetic Code Expansion; Non-Canonical Amino Acid; Photoactivation; Photocaged Glutamate; Protein Design; Protein Engineering; Protein Modifications.