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Chemical Tagging with tert ‐Butyl and Trimethylsilyl Groups for Measuring Intermolecular Nuclear Overhauser Effects in a Large Protein–Ligand Complex
Author(s) -
Jabar Shereen,
Adams Luke A.,
Wang Yao,
Aurelio Luigi,
Graham Bim,
Otting Gottfried
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.201703531
Subject(s) - nuclear overhauser effect , chemistry , intermolecular force , trimethylsilyl , dna , ligand (biochemistry) , cysteine , chemical shift , chemical modification , nuclear magnetic resonance spectroscopy , quantum chemical , stereochemistry , molecule , biochemistry , organic chemistry , receptor , enzyme
Intermolecular 1 H‐ 1 H nuclear Overhauser effects (NOE) present a powerful tool to assess contacts between proteins and binding partners, but are difficult to identify for complexes of high molecular weight. This report shows that intermolecular NOEs can readily be observed following chemical labeling with tert ‐butyl or trimethylsilyl (TMS) groups. Proteins can be furnished with tert ‐butyl or TMS groups site‐specifically using genetically encoded unnatural amino acids or by chemical modification of single cysteine residues. No isotope labeling is required. The approach is demonstrated with the 95 kDa complex between tetrameric E. coli single‐stranded DNA binding protein (SSB) and single‐stranded DNA.