Protein painting for structural and binding site analysis via intracellular lysine reactivity profiling with o-phthalaldehyde.

ABSTRACT: The three-dimensional structure and the molecular interaction of proteins determine their roles in many cellular processes. Chemical protein painting with protein mass spectrometry can identify changes in structural conformations and molecular interactions of proteins including their binding sites. Nevertheless, most current protein painting techniques identify protein targets and binding sites of drugs in vitro using a cell lysate or purified protein. Here, we tested 11 membrane-permeable lysine-reactive chemical probes for intracellular covalent labeling of endogenous proteins, which reveals ortho-phthalaldehyde (OPA) as the most reactive probe in the intracellular environment. An MS workflow and a new data analysis strategy termed RAPID (Reactive Amino acid Profiling by In

SUBMITTER: Zheng Z 

PROVIDER: S-EPMC11040650 | biostudies-literature | 2024 Apr

REPOSITORIES: biostudies-literature