Open AccessEngineering a methyllysine reader with photoactive amino acid in mammalian cells
Author(s) -
Simran Arora,
Sushma Sappa,
Kathryn Hinkelman,
Kabirul Islam
Publication year - 2020
Publication title -
chemical communications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.837
H-Index - 333
eISSN - 1364-548X
pISSN - 1359-7345
DOI - 10.1039/d0cc03814h
Subject(s) - chromodomain , interactome , chemistry , biochemistry , amino acid , biology , computational biology , gene , helicase , rna
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.