Repository logo

Infoscience

  • English
  • French
Log In
Logo EPFL, École polytechnique fédérale de Lausanne

Infoscience

  • English
  • French
Log In
  1. Home
  2. Academic and Research Output
  3. Journal articles
  4. Structure and backbone dynamics of a microcrystalline metalloprotein by solid-state NMR
 
research article

Structure and backbone dynamics of a microcrystalline metalloprotein by solid-state NMR

Knight, Michael J.
•
Pell, Andrew J.
•
Bertini, Ivano
Show more
2012
Proceedings Of The National Academy Of Sciences Of The United States Of America (PNAS)

We introduce a new approach to improve structural and dynamical determination of large metalloproteins using solid-state nuclear magnetic resonance (NMR) with H-1 detection under ultrafast magic angle spinning (MAS). The approach is based on the rapid and sensitive acquisition of an extensive set of N-15 and C-13 nuclear relaxation rates. The system on which we demonstrate these methods is the enzyme Cu, Zn superoxide dismutase (SOD), which coordinates a Cu ion available either in Cu+ (diamagnetic) or Cu2+ (paramagnetic) form. Paramagnetic relaxation enhancements are obtained from the difference in rates measured in the two forms and are employed as structural constraints for the determination of the protein structure. When added to H-1-H-1 distance restraints, they are shown to yield a twofold improvement of the precision of the structure. Site-specific order parameters and timescales of motion are obtained by a Gaussian axial fluctuation (GAF) analysis of the relaxation rates of the diamagnetic molecule, and interpreted in relation to backbone structure and metal binding. Timescales for motion are found to be in the range of the overall correlation time in solution, where internal motions characterized here would not be observable.

  • Details
  • Metrics
Type
research article
DOI
10.1073/pnas.1204515109
Web of Science ID

WOS:000306642100021

Author(s)
Knight, Michael J.
•
Pell, Andrew J.
•
Bertini, Ivano
•
Felli, Isabella C.
•
Gonnelli, Leonardo
•
Pierattelli, Roberta
•
Herrmann, Torsten
•
Emsley, Lyndon  
•
Pintacuda, Guido
Date Issued

2012

Publisher

National Academy of Sciences

Published in
Proceedings Of The National Academy Of Sciences Of The United States Of America (PNAS)
Volume

109

Issue

28

Start page

11095

End page

11100

Subjects

paramagnetism

•

nuclear relaxation rates

•

copper

•

microcrystal

Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
LRM  
Available on Infoscience
January 8, 2015
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/109987
Logo EPFL, École polytechnique fédérale de Lausanne
  • Contact
  • infoscience@epfl.ch

  • Follow us on Facebook
  • Follow us on Instagram
  • Follow us on LinkedIn
  • Follow us on X
  • Follow us on Youtube
AccessibilityLegal noticePrivacy policyCookie settingsEnd User AgreementGet helpFeedback

Infoscience is a service managed and provided by the Library and IT Services of EPFL. © EPFL, tous droits réservés