Dipolar chemical shift correlation spectroscopy for homonuclear carbon distance measurements in proteins in the solid state: Application to structure determination and refinement

Xiaohu Peng, David Libich, Rafal Janik, George Harauz, Vladimir Ladizhansky

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

High-resolution solid-state NMR spectroscopy has become a promising tool for protein structure determination. Here, we describe a new dipolar-chemical shift correlation experiment for the measurement of homonuclear 13C-13C distances in uniformly 13C, 15N-labeled proteins and demonstrate its suitability for protein structure determination and refinement. The experiments were carried out on the β1 immunoglobulin binding domain of protein G (GB1). Both intraresidue and interresidue distances between carbonyl atoms and atoms in the aliphatic side chains were collected using a three-dimensional chemical shift correlation spectroscopy experiment that uses homogeneously broadened rotational resonance recoupling for carbon mixing. A steady-state approximation for the polarization transfer function was employed in data analysis, and a total of 100 intramolecular distances were determined, all in the range 2.5-5.5 Å. An additional 41 dipolar contacts were detected, but the corresponding distances could not be accurately quantified. Additional distance and torsional restraints were derived from the proton-driven spin diffusion measurements and from the chemical shift analysis, respectively. Using all these restraints, it was possible to refine the structure of GB1 to a root-mean square deviation of 0.8 Å. The approach is of general applicability for peptides and small proteins and can be easily incorporated into a structure determination and refinement protocol.

Original languageEnglish (US)
Pages (from-to)359-369
Number of pages11
JournalJournal of the American Chemical Society
Volume130
Issue number1
DOIs
StatePublished - Jan 9 2008
Externally publishedYes

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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