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

Ivano Bertini, Anusarka Bhaumik, Gaël De Paëpe, Robert G. Griffin, Moreno Lelli, Józef R. Lewandowski, Claudio Luchinat

Research output: Contribution to journalArticlepeer-review

100 Scopus citations

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 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.

Original languageEnglish (US)
Pages (from-to)1032-1040
Number of pages9
JournalJournal of the American Chemical Society
Volume132
Issue number3
DOIs
StatePublished - 2010

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