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Gd3+-chelated lipid accelerates solid-state NMR spectroscopy of seven-transmembrane proteins

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Abstract

Solid-state NMR (SSNMR) is an attractive technique for studying large membrane proteins in membrane-mimetic environments. However, SSNMR experiments often suffer from low efficiency, due to the inherent low sensitivity and the long recycle delays needed to recover the magnetization. Here we demonstrate that the incorporation of a small amount of a Gd3+-chelated lipid, Gd3+-DMPE-DTPA, into proteoliposomes greatly shortens the spin–lattice relaxation time (1H-T 1) of lipid-reconstituted membrane proteins and accelerates the data collection. This effect has been evaluated on a 30 kDa, seven-transmembrane protein, Leptosphaeria rhodopsin. With the Gd3+-chelated lipid, we can perform 2D SSNMR experiments 3 times faster than by diamagnetic control. By combining this paramagnetic relaxation-assisted data collection with non-uniform sampling, the 3D experimental times are reduced eightfold with respect to traditional 3D experiments on diamagnetic samples. A comparison between the paramagnetic relaxation enhancement (PRE) effects of Cu2+- and Gd3+-chelated lipids indicates the much higher relaxivity of the latter. Hence, a tenfold lower concentration is needed for Gd3+-chelated lipids to achieve comparable PRE effects to Cu2+-chelated lipids. In addition, Gd3+-chelated lipids neither alter the protein structures nor induce significant line-width broadening of the protein signals. This work is expected to be beneficial for structural and dynamic studies of large membrane proteins by SSNMR.

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Acknowledgements

All NMR experiments were carried out at the Beijing NMR Center or the NMR Facility of the National Center for Protein Sciences at Peking University. This work was supported by the National Natural Science Foundation of China (Contract No. 31470727), by the National Key Research and Development Program from the Ministry of Science and Technology of the People’s Republic of China (Contract No. 2016YFA0501203), by the Beijing National Laboratory for Molecular Sciences, and by the start-up funds from Peking University. S.W. is a recipient of the 1000 plan for young talent program of China. We thank Prof. Leonid S. Brown, from University of Guelph, for providing LR expression colonies. We thank Dr. Yong Liang, from Xiamen University, for assistance in preparation of Gd3+-DOTA.

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  1. ShengQi Xiang

    Present address: NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands

Authors and Affiliations

  1. College of Chemistry and Molecular Engineering, Peking University, Yiheyuan Rd. 5th, Beijing, China

    Chang Liu, Jing Liu, Xiaojun Xu & Shenlin Wang

  2. Beijing NMR Center, Peking University, Yiheyuan Rd. 5th, Beijing, China

    Chang Liu, Jing Liu, Xiaojun Xu & Shenlin Wang

  3. Department NMR-Based Structural Biology, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077, Göttingen, Germany

    ShengQi Xiang

  4. National Laboratories of Beijing National Laboratory for Molecular Science, Beijing, China

    Shenlin Wang

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  1. Chang Liu
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Correspondence to Shenlin Wang.

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Liu, C., Liu, J., Xu, X. et al. Gd3+-chelated lipid accelerates solid-state NMR spectroscopy of seven-transmembrane proteins. J Biomol NMR 68, 203–214 (2017). https://doi.org/10.1007/s10858-017-0120-y

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