Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR

Nathaniel Traaseth, Gianluigi Veglia

Research output: Contribution to journalArticle

Abstract

We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR spectra of membrane proteins in fluid lipid membranes with broad lines and high redundancy in the primary sequence. We implemented this approach in both heteronuclear 15N- 13Cα and homonuclear 13C-13C dipolar assisted rotational resonance (DARR) correlation experiments. We demonstrate its efficacy for the membrane protein phospholamban reconstituted in fluid PC/PE/PA lipid bilayers. The main advantage of this method is to discriminate overlapped 13Cα resonances by strategically labeling the preceding residue. This method is highly complementary to 13Ci-1′-15Ni- 13Ciα and 13Ci-1α-15N i-1-13Ci′ experiments to distinguish inter-residue spin systems at a minimal cost to signal-to-noise.

Original languageEnglish (US)
Pages (from-to)18-24
Number of pages7
JournalJournal of Magnetic Resonance
Volume211
Issue number1
DOIs
StatePublished - Jul 2011

Fingerprint

Magic angle spinning
Labeling
metal spinning
marking
Membrane Proteins
Nuclear magnetic resonance
membranes
proteins
nuclear magnetic resonance
Lipid bilayers
Fluids
lipids
Membrane Lipids
Redundancy
fluids
Experiments
Lipid Bilayers
redundancy
Noise
costs

Keywords

  • Frequency-selective dipolar recoupling
  • Heteronuclear recoupling
  • Magic angle spinning
  • Membrane proteins
  • Phospholamban
  • REDOR
  • Solid-state NMR

ASJC Scopus subject areas

  • Nuclear and High Energy Physics
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics

Cite this

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abstract = "We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR spectra of membrane proteins in fluid lipid membranes with broad lines and high redundancy in the primary sequence. We implemented this approach in both heteronuclear 15N- 13Cα and homonuclear 13C-13C dipolar assisted rotational resonance (DARR) correlation experiments. We demonstrate its efficacy for the membrane protein phospholamban reconstituted in fluid PC/PE/PA lipid bilayers. The main advantage of this method is to discriminate overlapped 13Cα resonances by strategically labeling the preceding residue. This method is highly complementary to 13Ci-1′-15Ni- 13Ciα and 13Ci-1α-15N i-1-13Ci′ experiments to distinguish inter-residue spin systems at a minimal cost to signal-to-noise.",
keywords = "Frequency-selective dipolar recoupling, Heteronuclear recoupling, Magic angle spinning, Membrane proteins, Phospholamban, REDOR, Solid-state NMR",
author = "Nathaniel Traaseth and Gianluigi Veglia",
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AU - Traaseth, Nathaniel

AU - Veglia, Gianluigi

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N2 - We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR spectra of membrane proteins in fluid lipid membranes with broad lines and high redundancy in the primary sequence. We implemented this approach in both heteronuclear 15N- 13Cα and homonuclear 13C-13C dipolar assisted rotational resonance (DARR) correlation experiments. We demonstrate its efficacy for the membrane protein phospholamban reconstituted in fluid PC/PE/PA lipid bilayers. The main advantage of this method is to discriminate overlapped 13Cα resonances by strategically labeling the preceding residue. This method is highly complementary to 13Ci-1′-15Ni- 13Ciα and 13Ci-1α-15N i-1-13Ci′ experiments to distinguish inter-residue spin systems at a minimal cost to signal-to-noise.

AB - We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR spectra of membrane proteins in fluid lipid membranes with broad lines and high redundancy in the primary sequence. We implemented this approach in both heteronuclear 15N- 13Cα and homonuclear 13C-13C dipolar assisted rotational resonance (DARR) correlation experiments. We demonstrate its efficacy for the membrane protein phospholamban reconstituted in fluid PC/PE/PA lipid bilayers. The main advantage of this method is to discriminate overlapped 13Cα resonances by strategically labeling the preceding residue. This method is highly complementary to 13Ci-1′-15Ni- 13Ciα and 13Ci-1α-15N i-1-13Ci′ experiments to distinguish inter-residue spin systems at a minimal cost to signal-to-noise.

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KW - REDOR

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