3D S3 experiment

This is NYSBC standard data set and pulse program for S3 experiment - DARR-DCP-DCP-DARR. The experiment utilizes DNP enhancement to conduct multiple (3-4) polarization transfers. It can be used for sequential sidechain correlation. It can also be used as a 2D for sequential Calpha correlation. The pulse program S3.nysbc is located in the dataset directory.

Citation

Efficient assignment and NMR analysis of an intact virus using sequential side-chain correlations and DNP sensitization. Ivan V. Sergeyev, Boris Itin, Rivkah Rogawski, Loren A. Day, and Ann E. McDermott, PNAS, 2017 May 16; 114(20): 5171–5176. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5441803

Probe and power limitations

DCP experiments require long simultaneous medium to high power pulses on three channels simultaneously. Adjust the probe power length and safety limitations in the pulse sequence to follow your probe specifications. When in doubt, contact the manufacturer.

Protocol

Use very short first CP pulse to only transfer magnetization from 1H to Calpha. DCP conditions for N-C and C-N transfers are identical. This data set uses NUS (non uniform sampling) to improve resolution. Only use NUS if S/N > 10:1. For better resolution use longer acquisition times in the indirect dimensions (8-12ms for 13C and 5-7ms for 15N)

Samples

The experiments were performed on uniformly 13C/15N labeled amyloid and Pf1 bacteriophage samples using "DNP juice" of 60/30/10 d6-glycerol/D2O/H2O with 15mM Amupol concentration. 600MHz/395GHz DNP instrument.

Processing and analysis

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Pulse Sequence and Parameter Set