biostudies-literatureArora SNational Institutes of HealthNIGMS NIH HHSNational Science Foundation12210-12213https://www.ebi.ac.uk/biostudies/studies/S-EPMC7573786biostudies-literatureUnknown56(81)Methyllysine sites in proteins are recognized by an array of reader domains that mediate protein-protein interactions for controlling cellular processes. Herein, we engineer a chromodomain, an essential methyllysine reader, to carry 4-azido-l-phenylalanine (AzF) via amber suppressor mutagenesis and demonstrate its potential to bind and crosslink methylated proteins in human cells. We further develop a first-of-its kind chromodomain variant bearing two AzF units with enhanced crosslinking potential suitable for profiling the transient methyllysine interactome.Chemical communications (Cambridge, England)Engineering a methyllysine reader with photoactive amino acid in mammalian cells.PMC7573786R01GM123234R01 GM130752MCB-1817692R01 GM123234R01GM130752Hinkelman KIslam KArora SSappa SfalseEngineering a methyllysine reader with photoactive amino acid in mammalian cells.Methyllysine sites in proteins are recognized by an array of reader domains that mediate protein-protein interactions for controlling cellular processes. Herein, we engineer a chromodomain, an essential methyllysine reader, to carry 4-azido-l-phenylalanine (AzF) via amber suppressor mutagenesis and demonstrate its potential to bind and crosslink methylated proteins in human cells. We further develop a first-of-its kind chromodomain variant bearing two AzF units with enhanced crosslinking potential suitable for profiling the transient methyllysine interactome.2020-01-01T00:00:00Z2020 Oct2026-04-29T10:24:21.442Z2026-04-07T15:09:22.594ZS-EPMC75737863292602310.1039/d0cc03814h