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Chemical Tagging with tert ‐Butyl and Trimethylsilyl Groups for Measuring Intermolecular Nuclear Overhauser Effects in a Large Protein–Ligand Complex
- Source :
- Chemistry – A European Journal. 23:13033-13036
- Publication Year :
- 2017
- Publisher :
- Wiley, 2017.
-
Abstract
- Intermolecular 1H-1H 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. We show that intermolecular NOEs can readily be observed following chemical labeling with t-butyl or trimethylsilyl groups. Proteins can be furnished with t-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.
- Subjects :
- Models, Molecular
0301 basic medicine
Trimethylsilyl Compounds
Trimethylsilyl
Stereochemistry
DNA, Single-Stranded
Nuclear Overhauser effect
Ligands
010402 general chemistry
01 natural sciences
DNA-binding protein
Catalysis
Structure-Activity Relationship
03 medical and health sciences
chemistry.chemical_compound
Escherichia coli
Organic chemistry
Amino Acid Sequence
Nuclear Magnetic Resonance, Biomolecular
chemistry.chemical_classification
Binding Sites
Molecular Structure
Chemistry
Organic Chemistry
Intermolecular force
Chemical modification
General Chemistry
0104 chemical sciences
Amino acid
DNA-Binding Proteins
030104 developmental biology
Isotope Labeling
DNA
Protein Binding
Cysteine
Subjects
Details
- ISSN :
- 15213765 and 09476539
- Volume :
- 23
- Database :
- OpenAIRE
- Journal :
- Chemistry – A European Journal
- Accession number :
- edsair.doi.dedup.....405c95c3479c9b827f7697d45892025a