Selective 1H-1H Distance Restraints in Fully Protonated Proteins by Very Fast Magic-Angle Spinning Solid-State NMR

Mukul G. Jain, Daniela Lalli, Jan Stanek, Chandrakala Gowda, Satya Prakash, Tom S. Schwarzer, Tobias Schubeis, Kathrin Castiglione, Loren B. Andreas, P. K. Madhu, Guido Pintacuda, Vipin Agarwal

Research output: Contribution to journalArticlepeer-review

Abstract

Very fast magic-angle spinning (MAS > 80 kHz) NMR combined with high-field magnets has enabled the acquisition of proton-detected spectra in fully protonated solid samples with sufficient resolution and sensitivity. One of the primary challenges in structure determination of protein is observing long-range 1H-1H contacts. Here we use band-selective spin-lock pulses to obtain selective 1H-1H contacts (e.g., HN-HN) on the order of 5-6 Å in fully protonated proteins at 111 kHz MAS. This approach is a major advancement in structural characterization of proteins given that magnetization can be selectively transferred between protons that are 5-6 Å apart despite the presence of other protons at shorter distance. The observed contacts are similar to those previously observed only in perdeuterated proteins with selective protonation. Simulations and experiments show the proposed method has performance that is superior to that of the currently used methods. The method is demonstrated on GB1 and a β-barrel membrane protein, AlkL.

Original languageEnglish
Pages (from-to)2399-2405
Number of pages7
JournalJournal of Physical Chemistry Letters
Volume8
Issue number11
DOIs
Publication statusPublished - 1 Jun 2017
Externally publishedYes

Fingerprint

Dive into the research topics of 'Selective 1H-1H Distance Restraints in Fully Protonated Proteins by Very Fast Magic-Angle Spinning Solid-State NMR'. Together they form a unique fingerprint.

Cite this