Pseudocontact shifts (PCSs) arise in para-magnetic 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 H-1-detected solid-state correlations provides facile and rapid detection and assignment of site-specific PCSs, including resolved H-1 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 H-1-H-1 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.

Rapid Measurement of Pseudocontact Shifts in Metalloproteins by Proton-Detected Solid-State NMR Spectroscopy / M.J. Knight; I.C. Felli; R. Pierattelli; I. Bertini; L. Emsley; T. Herrmann; G. Pintacuda. - In: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. - ISSN 0002-7863. - STAMPA. - 134:(2012), pp. 14730-14733. [10.1021/ja306813j]

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

FELLI, ISABELLA CATERINA;PIERATTELLI, ROBERTA;BERTINI, IVANO;
2012

Abstract

Pseudocontact shifts (PCSs) arise in para-magnetic 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 H-1-detected solid-state correlations provides facile and rapid detection and assignment of site-specific PCSs, including resolved H-1 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 H-1-H-1 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.
2012
134
14730
14733
M.J. Knight; I.C. Felli; R. Pierattelli; I. Bertini; L. Emsley; T. Herrmann; G. Pintacuda
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/753725
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