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Engineering a methyllysine reader with photoactive amino acid in mammalian cells.


ABSTRACT: 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.

SUBMITTER: Arora S 

PROVIDER: S-EPMC7573786 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Engineering a methyllysine reader with photoactive amino acid in mammalian cells.

Arora Simran S   Sappa Sushma S   Hinkelman Kathryn K   Islam Kabirul K  

Chemical communications (Cambridge, England) 20200914 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 potenti  ...[more]

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