Faculty Collaboration Database

¹H-detected MAS solid-state NMR experiments enable the simultaneous mapping of rigid and dynamic domains of membrane proteins. J Magn Reson 2017 Dec;285:101-107

Date

11/28/2017

Pubmed ID

29173803

Pubmed Central ID

PMC5764182

DOI

10.1016/j.jmr.2017.09.003

Scopus ID

2-s2.0-85034835475 (requires institutional sign-in at Scopus site)   15 Citations

Abstract

Magic angle spinning (MAS) solid-state NMR (ssNMR) spectroscopy is emerging as a unique method for the atomic resolution structure determination of native membrane proteins in lipid bilayers. Although ¹³C-detected ssNMR experiments continue to play a major role, recent technological developments have made it possible to carry out ¹H-detected experiments, boosting both sensitivity and resolution. Here, we describe a new set of ¹H-detected hybrid pulse sequences that combine through-bond and through-space correlation elements into single experiments, enabling the simultaneous detection of rigid and dynamic domains of membrane proteins. As proof-of-principle, we applied these new pulse sequences to the membrane protein phospholamban (PLN) reconstituted in lipid bilayers under moderate MAS conditions. The cross-polarization (CP) based elements enabled the detection of the relatively immobile residues of PLN in the transmembrane domain using through-space correlations; whereas the most dynamic region, which is in equilibrium between folded and unfolded states, was mapped by through-bond INEPT-based elements. These new ¹H-detected experiments will enable one to detect not only the most populated (ground) states of biomacromolecules, but also sparsely populated high-energy (excited) states for a complete characterization of protein free energy landscapes.

Author List

Gopinath T, Nelson SED, Veglia G

Author

Gopinath Tata PhD Assistant Professor in the Biophysics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Calcium-Binding Proteins
Carbon Isotopes
Lipid Bilayers
Membrane Proteins
Nitrogen Isotopes
Nuclear Magnetic Resonance, Biomolecular
Sensitivity and Specificity