Faculty Collaboration Database

Application of paramagnetic relaxation enhancements to accelerate the acquisition of 2D and 3D solid-state NMR spectra of oriented membrane proteins. Methods 2018 Apr 01;138-139:54-61

Date

12/25/2017

Pubmed ID

29274874

Pubmed Central ID

PMC5984106

DOI

10.1016/j.ymeth.2017.12.017

Scopus ID

2-s2.0-85044638879 (requires institutional sign-in at Scopus site)   15 Citations

Abstract

Oriented sample solid-state NMR (OS-ssNMR) spectroscopy is uniquely suited to determine membrane protein topology at the atomic resolution in liquid crystalline bilayers under physiological temperature. However, the inherent low sensitivity of this technique has hindered the throughput of multidimensional experiments necessary for resonance assignments and structure determination. In this work, we show that doping membrane protein bicelle preparations with paramagnetic ion chelated lipids and exploiting paramagnetic relaxation effects it is possible to accelerate the acquisition of both 2D and 3D multidimensional experiments with significant saving in time. We demonstrate the efficacy of this method for a small membrane protein, sarcolipin, reconstituted in DMPC/POPC/DHPC oriented bicelles. In particular, using Cu²⁺-DMPE-DTPA as a dopant, we observed a decrease of ¹H T₁ of sarcolipin by 2/3, allowing us to reduce the recycle delay up to 3 times. We anticipate that these new developments will enable the routine acquisition of multidimensional OS-ssNMR experiments.

Author List

Wang S, Gopinath T, 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

Animals
Humans
Membrane Proteins
Muscle Proteins
Nuclear Magnetic Resonance, Biomolecular
Protein Conformation
Proteolipids