experiment for 1H observe hCANH at fast MAS.

This is a 1H-13Calpha-15N crosscorrelation 3D experiment optimized for fast MAS (1.3mm probe at 60kHz spinning frequency).

Citation

https://www.ncbi.nlm.nih.gov/pubmed/25102442 E. Barbet-Massin, A.J. Pell, J.S. Retel, et al., Rapid Proton-Detected NMR Assignment for Proteins with Fast Magic Angle Spinning., JACS 136 (35), pp 12489-12497 (2014).

Sample heating

Frictional heating during ultra fast spinning may heat up a sample by up to 40K.

Citation

[https://www.biorxiv.org/content/10.1101/566729v1.full] TmDOTP : An NMR- based Thermometer for Magic Angle Spinning NMR Experiments Dongyu Zhang, Boris Itin, Ann E. McDermott; preprinted

Protocol

Use CPHSQC experiments to optimize matching conditions and spin lock length for HC and HN transfers and to calibrate 13C and 15N pulses. Use 175us Q3.1000 selective pulses for Calpha and C(O) inversion. Calculate power levels for the selective pulses using Bruker shapetool or edprosol routine. Experimentally calibrate the selective pulses for maximum signal. For DCP condition, use 25kHz/35kHz match with tangential shaped pulse.

Samples

The experiments were performed with a precipitated microcrystalline u-13C,15N labeled protein

Processing and analysis

Fourier transform a 2D experiment. Gaussian apodization is recommended.

Pulse Sequence and Parameter Set