NMR identification of protein surfaces using paramagnetic probes
- SmithKline Beecham Pharmaceuticals, Prussia, PA (USA)
Paramagnetic agents produce line broadening and thus cancellation of anti phase cross-peak components in two-dimensional correlated nulcear magnetic resonance spectra. The specificity of this effect was examined to determine its utility for identifying surface residues of proteins. Ubiquitin and hen egg white lysozyme, for which X-ray crystal structures and proton NMR assignments are available, served as test cases. Two relaxation reagents were employed, 4-hydroxy-2,2,6,6-tetramethylpiperidinyl-1 oxy and the gadolinium(III) diethylenetriaminepentaacetate complex ion. Correlations were sought between reagent-produced decreases of side-chain cross-peak volumes in double-quantum-filtered proton correlation (DQF-COSY) spectra and the solvent-exposed side-chain surface area of the corresponding residues. The lanthanide complex produced strong effects ascribable to association with carboxylate groups but was not otherwise useful in delineating surface residues. The nitroxyl, on the other hand, produced clear distinctions among the Val, Leu, and Ile residues that generally paralleled side-chain exposure in the crystal, although consistent correlations were not observed with residues of other types. Although an instance of possible specific protein-nitroxyl association was noted, the nitroxyl appears to be a tool for identifying hydrophobic surface residues.
- OSTI ID:
- 5736665
- Journal Information:
- Biochemistry; (USA), Vol. 29:43; ISSN 0006-2960
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GADOLINIUM COMPLEXES
PARAMAGNETISM
LYSOZYME
NUCLEAR MAGNETIC RESONANCE
DTPA
MOLECULAR STRUCTURE
NMR SPECTRA
OVERHAUSER EFFECT
RADICALS
SHAPE
UBIQUINONE
AMINO ACIDS
AROMATICS
BENZOQUINONES
CARBOXYLIC ACIDS
CHELATING AGENTS
COENZYMES
COMPLEXES
DRUGS
ENZYMES
GLYCOSYL HYDROLASES
HYDROLASES
MAGNETIC RESONANCE
MAGNETISM
O-GLYCOSYL HYDROLASES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
QUINONES
RADIOPROTECTIVE SUBSTANCES
RARE EARTH COMPLEXES
RESONANCE
SPECTRA
550201* - Biochemistry- Tracer Techniques