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 A, and is in good agreement with the X-ray structure (rmsd to X-ray 1.3 A). 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.

High-resolution solid-state NMR structure of a 17.6 kDa protein / I.Bertini; A.Bhaumik; G.De Paëpe; R.G.Griffin; M.Lelli; J.R.Lewandowski; C.Luchinat. - In: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. - ISSN 0002-7863. - STAMPA. - 132(3)(2010), pp. 1032-1040. [10.1021/ja906426p]

High-resolution solid-state NMR structure of a 17.6 kDa protein

BERTINI, IVANO;BHAUMIK, ANUSARKA;LELLI, MORENO
Writing – Original Draft Preparation
;
LUCHINAT, CLAUDIO
2010

Abstract

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 A, and is in good agreement with the X-ray structure (rmsd to X-ray 1.3 A). 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.
132(3)
1032
1040
Goal 3: Good health and well-being for people
I.Bertini; A.Bhaumik; G.De Paëpe; R.G.Griffin; M.Lelli; J.R.Lewandowski; C.Luchinat
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2158/371104
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