Direct amide N-15 to C-13 transfers for solid-state assignment experiments in deuterated proteins
Author / Producer
Date
2018-10
Publication Type
Journal Article
ETH Bibliography
yes
Citations
Altmetric
Data
Rights / License
Abstract
The assignment of protein backbone and side-chain NMR chemical shifts is the first step towards the characterization of protein structure. The recent introduction of proton detection in combination with fast MAS has opened up novel opportunities for assignment experiments. However, typical 3D sequential-assignment experiments using proton detection under fast MAS lead to signal intensities much smaller than the theoretically expected ones due to the low transfer efficiency of some of the steps. Here, we present a selective 3D experiment for deuterated and (amide) proton back-exchanged proteins where polarization is directly transferred from backbone nitrogen to selected backbone or sidechain carbons. The proposed pulse sequence uses only ¹H–¹⁵N cross-polarization (CP) transfers, which are, for deuterated proteins, about 30% more efficient than ¹H–¹³C CP transfers, and employs a dipolar version of the INEPT experiment for N–C transfer. By avoiding Hᴺ–C (Hᴺ stands for amide protons) and C–C CP transfers, we could achieve higher selectivity and increased signal intensities compared to other pulse sequences containing long-range CP transfers. The REDOR transfer is designed with an additional selective π pulse, which enables the selective transfer of the polarization to the desired ¹³C spins.
Permanent link
Publication status
published
Editor
Book title
Journal / series
Volume
72 (1)
Pages / Article No.
69 - 78
Publisher
Springer
Event
Edition / version
Methods
Software
Geographic location
Date collected
Date created
Subject
Solid-state NMR; MAS; Proteins; Magnetisation transfer
Organisational unit
03496 - Meier, Beat H. (emeritus) / Meier, Beat H. (emeritus)
08829 - Ernst, Matthias (Tit.-Prof.)
Notes
It was possible to publish this article open access thanks to a Swiss National Licence with the publisher.
Funding
159707 - NMR studies in the Solid State (SNF)
146757 - NMR studies in the Solid State (SNF)
741863 - Faster magic-angle spinning leads to a resolution revolution in biological solid-state NMR (EC)
146757 - NMR studies in the Solid State (SNF)
741863 - Faster magic-angle spinning leads to a resolution revolution in biological solid-state NMR (EC)