13C- and 1H-detection under fast MAS for the study of poorly available proteins: application to sub-milligram quantities of a 7 trans-membrane protein

Hugh R W Dannatt, Garrick F. Taylor, Krisztina Varga, Victoria A. Higman, Marc Philipp Pfeil, Lubica Asilmovska, Peter J. Judge, Anthony Watts*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

14 Citations (Scopus)

Abstract

We demonstrate that 13C-detected spectra recorded using fast (60 kHz) magic angle spinning on sub-milligram (<10 μmol) quantities of a protonated 7 trans-membrane helix protein (bacteriorhodopsin) in its native lipid environment are comparable in sensitivity and resolution to those recorded using 15-fold larger sample volumes with conventional solid state NMR methodology. We demonstrate the utility of proton-detected measurements which yield narrow 1H linewidths under these conditions, and that no structural alterations are observed. We propose that these methods will prove useful to gain structural information on membrane proteins with poor availability, which can be studied in their native lipid environments.

Original languageEnglish
Pages (from-to)17-23
JournalJournal of Biomolecular NMR
Volume62
Early online date21 Feb 2015
DOIs
Publication statusPublished - 21 Feb 2015

Keywords

  • C-detection
  • 7 trans-membrane proteins
  • Fast magic angle spinning
  • Heteronuclear detection
  • Low sample volumes
  • Poorly available proteins

Fingerprint

Dive into the research topics of '<sup>13</sup>C- and <sup>1</sup>H-detection under fast MAS for the study of poorly available proteins: application to sub-milligram quantities of a 7 trans-membrane protein'. Together they form a unique fingerprint.

Cite this