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

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.

SUBMITTER: Bertini I

PROVIDER: S-EPMC4437541 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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

Bertini Ivano I Bhaumik Anusarka A De Paëpe Gaël G Griffin Robert G RG Lelli Moreno M Lewandowski Józef R JR Luchinat Claudio C

Journal of the American Chemical Society 20100101 3

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(I ...[more]

PMID: 20041641

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