Paramagnetic shifts in solid-state NMR of proteins to elicit structural information

Proc Natl Acad Sci U S A. 2008 Nov 11;105(45):17284-9. doi: 10.1073/pnas.0708460105. Epub 2008 Nov 6.


The recent observation of pseudocontact shifts (pcs) in (13)C high-resolution solid-state NMR of paramagnetic proteins opens the way to their application as structural restraints. Here, by investigating a microcrystalline sample of cobalt(II)-substituted matrix metalloproteinase 12 [CoMMP-12 (159 AA, 17.5 kDa)], it is shown that a combined strategy of protein labeling and dilution of the paramagnetic species (i.e., (13)C-,(15)N-labeled CoMMP-12 diluted in unlabeled ZnMMP-12, and (13)C-,(15)N-labeled ZnMMP-12 diluted in unlabeled CoMMP-12) allows one to easily separate the pcs contributions originated from the protein internal metal (intramolecular pcs) from those due to the metals in neighboring proteins in the crystal lattice (intermolecular pcs) and that both can be used for structural purposes. It is demonstrated that intramolecular pcs are significant structural restraints helpful in increasing both precision and accuracy of the structure, which is a need in solid-state structural biology nowadays. Furthermore, intermolecular pcs provide unique information on positions and orientations of neighboring protein molecules in the solid phase.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cobalt
  • Isotope Labeling
  • Magnetics*
  • Matrix Metalloproteinase 12 / chemistry*
  • Models, Molecular*
  • Nuclear Magnetic Resonance, Biomolecular


  • Cobalt
  • Matrix Metalloproteinase 12

Associated data

  • PDB/2K9C