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

Mukul G Jain (1) , Daniela Lalli (2) , Stanek Jan (2) , Chandrakala Gowda (1) , Satya Prakash (1) , Tom S Schwarzer (3) , Tobias Schubeis (2) , Kathrin Castiglione (3) , Loren B Andreas (4) , P. K. Madhu (1, 5) , Guido Pintacuda (2) , Vipin Agarwal (1)

1 TCIS - TIFR Centre for Interdisciplinary Sciences [Hyderabad]
2 Biological Solid-State NMR Methods - Méthodes de RMN à l'état solide en biologie
3 Inst Biochem Engn
4 MPI-BPC - Max Planck Institute for Biophysical Chemistry
5 DCS - Department of Chemical Sciences [TIFR]

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 H-1-H-1 contacts. Here we use band-selective spin-lock pulses to obtain selective H-1-H-1 contacts (e.g., H-N-H-N) on the order of 5-6 angstrom 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 angstrom 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 beta-barrel membrane protein, AlkL.

Mots clés: structure determination; structural characterization; protein; GB1; beta-barrel membrane protein; AlkL; NMR; MAS

Domaines: Chimie analytique; Chimie théorique et/ou physique; Biochimie [q-bio.BM]

Mukul G Jain, Daniela Lalli, Stanek Jan, Chandrakala Gowda, Satya Prakash, et al.. Selective 1H–1H Distance Restraints in Fully Protonated Proteins by Very Fast Magic-Angle Spinning Solid-State NMR. Journal of Physical Chemistry Letters, 2017, 8 (11), pp.2399-2405. ⟨10.1021/acs.jpclett.7b00983⟩. ⟨hal-01544734⟩