Abstract
Proton-detected solid-state NMR (ssNMR) spectroscopy has dramatically improved the sensitivity and resolution of fast magic angle spinning (MAS) methods. While relatively straightforward for fibers and crystalline samples, the routine application of these techniques to membrane protein samples is still challenging. This is due to the low sensitivity of these samples, which require high lipid:protein ratios to maintain the structural and functional integrity of membrane proteins. We previously introduced a family of novel polarization optimized experiments (POE) that enable to make the best of nuclear polarization and obtain multiple-acquisitions from a single pulse sequence and one receiver. Here, we present the 1H-detected versions of POE using ultrafast MAS ssNMR. Specifically, we implemented proton detection into our three main POE strategies, H-DUMAS, H-MEIOSIS, and H-MAeSTOSO, achieving the acquisition of up to ten different experiments using a single pulse sequence. We tested these experiments on a model compound N-Acetyl-Val-Leu dipeptide and applied to a six transmembrane acetate transporter, SatP, reconstituted in lipid membranes. These new methods will speed up the spectroscopy of challenging biomacromolecules such as membrane proteins.
Original language | English (US) |
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Article number | 106664 |
Journal | Journal of Magnetic Resonance |
Volume | 310 |
DOIs | |
State | Published - Jan 2020 |
Bibliographical note
Funding Information:The National Institute of Health (GM 64742, HL 144130, 1S10OD021536 to G.V) supported this work. All the experiments were carried out at the Minnesota NMR Center.
Publisher Copyright:
© 2019 Elsevier Inc.
Keywords
- H-DUMAS
- H-MAeSTOSO
- H-MEIOSIS
- Membrane proteins
- Multi-acquisition
- Polarization optimized experiments (POE)
- SIM-CP
- Solid-state NMR
- Ultra-fast magic angle spinning