Genetic Encoding of a Trifunctional Photo-Cross-Linker with a Cleavable Alkyl Ester Moiety.

ABSTRACT: Genetically encoded photo-cross-linkable amino acids (PAAs) are powerful tools for analyzing direct protein-protein interactions (PPIs) in mammalian cells. Cleavable PAAs are particularly useful, enabling covalent capture and subsequent release of interacting partners, which facilitates the characterization of interaction interfaces using mass spectrometry. However, the limited options for cleavable linker structures have restricted the design of PAAs. In this study, we genetically encoded a novel trifunctional PAA, DiZAAsu, which contains three distinct chemical groups: diazirine, alkyne, and alkaline-cleavable alkyl ester moieties. An archaeal pyrrolysyl-tRNA synthetase was engineered to incorporate DiZAAsu efficiently into proteins in mammalian cells. We demonstrated the in-cell photore

SUBMITTER: Takayama M 

PROVIDER: S-EPMC12859177 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature