HRMAS: DIPSY-13C 3D in u-13C,15N KcsA

HCCH-TOCSY 3D sequence with inverse correlation using multiple inept transfer and C-C DIPSI3 spinlock

Citation

L.E. Kay, G.Y. Xu, A.U. Singer, D.R. Muhandiram & J. D. Forman-Kay, J. Magn. Reson. B 101, 333 - 337 (1993) https://www.sciencedirect.com/science/article/pii/S1064186683710538?via%3Dihub

Protocol

a. This sequence works well in 1H solvents. b. Optimize the number of DIPSI loops. More loops show more connectivity but also produce more sample heating and cause more signal loss. c. DIPSI/TOCSY spinlock has to be rotor synchronized. RF = SF*1 or 0.5 otherwise it is unphaseable. d. Decrease trim pulse to 1ms

Samples

The experiments were performed with a precipitated U-13C/15N KcsA, LPR =1, 9:1 DOPE-DOPS D2O citrate buffer pH 7.5

Processing and analysis

Pulse Sequence and Parameter Set