Premium
Covalent Capturing of Transient SUMO–SIM Interactions Using Unnatural Amino Acid Mutagenesis and Photocrosslinking
Author(s) -
Taupitz Kim F.,
Dörner Wolfgang,
Mootz Henning D.
Publication year - 2017
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201605619
Subject(s) - covalent bond , amino acid , mutagenesis , chemistry , amino acid residue , protein–protein interaction , ubiquitin , sumo protein , computational biology , biochemistry , biophysics , combinatorial chemistry , stereochemistry , peptide sequence , biology , mutation , organic chemistry , gene
The weak interaction between the post‐translational modifier SUMO (small ubiquitin‐like modifier) and proteins containing the SUMO‐interacting motif (SIM) poses limitations to the identification of interaction partners of SUMOylated proteins and to the mapping of the interfaces. To overcome these limitations, genetically encoded photocrosslinker amino acids were incorporated close to the SIM‐interaction groove in human SUMO1. UV irradiation resulted in the desired covalent crosslinks both in a purified protein environment and in cell extracts.