Efficiency of Double Cross Polarization in Magic-Angle Spinning Solid-State NMR Studies on Membrane Proteins

doi:10.11938/cjmr20162528

Abstract

Abstract: The efficiency of multiple-dimensional heteronuclear correlation experiments in membrane protein studies is partially determined by the efficiency of double cross polarization (DCP). DCP is a basic block in the pulse sequences to probe the connections between the ¹⁵N and ¹³C nuclei. In this study, three membrane proteins were studied experimentally to determine how the dynamics properties of the proteins affect the DCP efficiency in solid-state heteronuclear correlation experiments. Under the same experimental conditions, the AQPZ protein showed the highest DCP efficiency (31%), while the EV71 2B protein had the lowest efficiency (14%) and the DAGK protein in between (23%). Longitudinal lattice relaxation time in the rotating frame (T_1ρ) and dipole coupling constant (D_HN) were also measured for these proteins. It was found that the DCP efficiency depended strongly on T_1ρ, but only weakly on D_HN. Based on the experimental data, a model to correlate the T_1ρ of protein to the DCP efficiency was established, with which the DCP efficiency could be predicted by measuring the T_1ρ of membrane proteins.

Key words: solid-state NMR, magic-angle spinning (MAS), membrane protein, double cross polarization (DCP), T_1ρ

CLC Number:

O482.53

SUN Yi, CHEN Yan-ke, LI Jian-ping, ZHAO Yong-xiang, YANG Jun. Efficiency of Double Cross Polarization in Magic-Angle Spinning Solid-State NMR Studies on Membrane Proteins[J]. Chinese Journal of Magnetic Resonance, 2017, 34(3): 257-265.

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