Rapid Measurement of Pseudocontact Shifts in Metalloproteins by Proton-Detected Solid-State NMR Spectroscopy

Pseudocontact shifts (PCSs) arise in paramagnetic systems in which the susceptibility tensor is anisotropic. PCSs depend upon the distance from the paramagnetic center and the position relative to the susceptibility tensor, and they can be used as structural restraints in protein structure determina...

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Published inJournal of the American Chemical Society Vol. 134; no. 36; pp. 14730 - 14733
Main Authors Knight, Michael J, Felli, Isabella C, Pierattelli, Roberta, Bertini, Ivano, Emsley, Lyndon, Herrmann, Torsten, Pintacuda, Guido
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 12.09.2012
Subjects
Analytical chemistry
Chemical Sciences
Cobalt - chemistry
Magnetic Resonance Spectroscopy - standards
Metalloproteins - chemistry
Models, Molecular
or physical chemistry
Protons
Reference Standards
Superoxide Dismutase - chemistry
Superoxide Dismutase - metabolism
Theoretical and
DOMAIN
PROTEIN
COMPLEXES
ENHANCEMENT
PARAMAGNETIC METALLOPROTEINS
DYNAMICS
ASSIGNMENT
RELAXATION
IONS
SUPEROXIDE-DISMUTASE
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Summary:Pseudocontact shifts (PCSs) arise in paramagnetic systems in which the susceptibility tensor is anisotropic. PCSs depend upon the distance from the paramagnetic center and the position relative to the susceptibility tensor, and they can be used as structural restraints in protein structure determination. We show that the use of 1H-detected solid-state correlations provides facile and rapid detection and assignment of site-specific PCSs, including resolved 1H PCSs, in a large metalloprotein, Co2+-substituted superoxide dismutase (Co2+-SOD). With only 3 mg of sample and a small set of experiments, several hundred PCSs were measured and assigned, and these PCSs were subsequently used in combination with 1H–1H distance and dihedral angle restraints to determine the protein backbone geometry with a precision paralleling those of state-of-the-art liquid-state determinations of diamagnetic proteins, including a well-defined active site.
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ISSN:0002-7863
1520-5126
DOI:10.1021/ja306813j