TY - JOUR
T1 - High-resolution solid-state NMR structure of a 17.6 kDa protein
AU - Bertini, Ivano
AU - Bhaumik, Anusarka
AU - De Paëpe, Gaël
AU - Griffin, Robert G.
AU - Lelli, Moreno
AU - Lewandowski, Józef R.
AU - Luchinat, Claudio
PY - 2010
Y1 - 2010
N2 - The use of pseudocontact shifts arising from paramagnetic metal ions in a microcrystalline protein sample is proposed as a strategy to obtain unambiguous signal assignments in solid-state NMR spectra enabling distance extraction for protein structure calculation. With this strategy, 777 unambiguous (281 sequential, 217 medium-range, and 279 long-range) distance restraints could be obtained from PDSD, DARR, CHHC, and the recently introduced PAR and PAIN-CP solid-state experiments for the cobalt(II)-substituted catalytic domain of matrix metalloproteinase 12 (159 amino acids, 17.6 kDa). The obtained structure is a high resolution one, with backbone rmsd of 1.0 ± 0.2 Å, and is in good agreement with the X-ray structure (rmsd to X-ray 1.3 Å). The proposed strategy, which may be generalized for nonmetalloproteins with the use of paramagnetic tags, represents a significant step ahead in protein structure determination using solid-state NMR.
AB - The use of pseudocontact shifts arising from paramagnetic metal ions in a microcrystalline protein sample is proposed as a strategy to obtain unambiguous signal assignments in solid-state NMR spectra enabling distance extraction for protein structure calculation. With this strategy, 777 unambiguous (281 sequential, 217 medium-range, and 279 long-range) distance restraints could be obtained from PDSD, DARR, CHHC, and the recently introduced PAR and PAIN-CP solid-state experiments for the cobalt(II)-substituted catalytic domain of matrix metalloproteinase 12 (159 amino acids, 17.6 kDa). The obtained structure is a high resolution one, with backbone rmsd of 1.0 ± 0.2 Å, and is in good agreement with the X-ray structure (rmsd to X-ray 1.3 Å). The proposed strategy, which may be generalized for nonmetalloproteins with the use of paramagnetic tags, represents a significant step ahead in protein structure determination using solid-state NMR.
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U2 - 10.1021/ja906426p
DO - 10.1021/ja906426p
M3 - Article
C2 - 20041641
AN - SCOPUS:76149116565
SN - 0002-7863
VL - 132
SP - 1032
EP - 1040
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 3
ER -