Spectral fitting for signal assignment and structural analysis of uniformly 13C-labeled solid proteins by simulated annealing based on chemical shifts and spin dynamics

J Biomol NMR. 2007 Aug;38(4):325-39. doi: 10.1007/s10858-007-9170-x. Epub 2007 Jul 6.

Abstract

We describe an approach for the signal assignment and structural analysis with a suite of two-dimensional (13)C-(13)C magic-angle-spinning solid-state NMR spectra of uniformly (13)C-labeled peptides and proteins. We directly fit the calculated spectra to experimental ones by simulated annealing in restrained molecular dynamics program CNS as a function of atomic coordinates. The spectra are calculated from the conformation dependent chemical shift obtained with SHIFTX and the cross-peak intensities computed for recoupled dipolar interactions. This method was applied to a membrane-bound 14-residue peptide, mastoparan-X. The obtained C', C(alpha) and C(beta) chemical shifts agreed with those reported previously at the precisions of 0.2, 0.7 and 0.4 ppm, respectively. This spectral fitting program also provides backbone dihedral angles with a precision of about 50 degrees from the spectra even with resonance overlaps. The restraints on the angles were improved by applying protein database program TALOS to the obtained chemical shifts. The peptide structure provided by these restraints was consistent with the reported structure at the backbone RMSD of about 1 A.

MeSH terms

  • Carbon Isotopes / chemistry
  • Computational Biology / methods
  • Intercellular Signaling Peptides and Proteins
  • Models, Molecular
  • Models, Statistical
  • Molecular Conformation
  • Nitrogen Isotopes / chemistry
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Peptides / chemistry*
  • Protein Conformation
  • Protein Structure, Secondary
  • Proteins / chemistry
  • Software
  • Time Factors

Substances

  • Carbon Isotopes
  • Intercellular Signaling Peptides and Proteins
  • Nitrogen Isotopes
  • Peptides
  • Proteins
  • mastoparan X