Site-specific dual encoding and labeling of proteins via genetic code expansion

• Published in: Cell chemical biology Vol. 30; no. 4; pp. 343 - 361
• Main Authors: Bednar, Riley M., Karplus, P. Andrew, Mehl, Ryan A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Ltd 20.04.2023
• Subjects: Amino Acids - chemistry; bioorthogonal chemistry; DEAL; dual encoding and labeling; Genetic Code; genetic code expansion; protein bioconjugation; protein engineering; Proteins - metabolism; dual encoding and labeling; bioorthogonal chemistry; protein engineering; genetic code expansion; DEAL; protein bioconjugation
• ISSN: 2451-9456; 2451-9448; 2451-9456
• DOI: 10.1016/j.chembiol.2023.03.004

The ability to selectively modify proteins at two or more defined locations opens new avenues for manipulating, engineering, and studying living systems. As a chemical biology tool for the site-specific encoding of non-canonical amino acids into proteins in vivo, genetic code expansion (GCE) represents a powerful tool to achieve such modifications with minimal disruption to structure and function through a two-step “dual encoding and labeling” (DEAL) process. In this review, we summarize the state of the field of DEAL using GCE. In doing so, we describe the basic principles of GCE-based DEAL, catalog compatible encoding systems and reactions, explore demonstrated and potential applications, highlight emerging paradigms in DEAL methodologies, and propose novel solutions to current limitations. [Display omitted] Site-specific dual labeling of proteins with genetic code expansion is a sought-after ability for studying and engineering proteins. In this review, Bednar et al. provide a conceptual framework and road map for navigating this powerful chemical biology tool.