Developing a DeSipher method for conformational change detection of membrane receptor complex at residue resolution

Liu, Q.; He, Q.-t.; Lyu, X.-x.; Yang, F.; Zhu, Z.-l.; Xiao, P.; Yang, Z.; Zhang, F.; Yang, Z.-y.; Wang, X.-y.; Sun, P.; Wang, Q.-w.; Qu, C.-x.; Gong, Z.; Lin, J.-Y.; Xu, Z.; Song, S.-l.; Huang, S.-m.; Guo, S.-c.; Han, M.-j.; Zhu, K.-k.; Chen, X.; Kahsai, A. W.; Xiao, K.-H.; Kong, W.; Yu, X.; Ruan, K.; Li, F.-h.; Niu, X.-g.; Jin, C.-w.; Wang, J.; Sun, J.

doi:10.1101/2019.12.18.873729

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bioRxiv · 10.1101/2019.12.18.873729

Developing a DeSipher method for conformational change detection of membrane receptor complex at residue resolution

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Abstract

While one dimensional 1H nuclear magnetic resonance (1D 1H-NMR) spectroscopy is one of the most important and convenient method for measuring conformation change in biomacromolecules, characterization of protein dynamics in large membrane protein complexes by 1D 1H-NMR remains challenging, due to the difficulty of spectra assignment, low signal-to-noise ratio (S/N) and the need for large amount of protein. Here we report the site-specific incorporation of 4-trimethylsilyl phenylalanine (TMSiPhe) into proteins, through genetic code expansion in Escherichia coli cells, and the measurement of multiple conformational states in membrane protein complex by 1D 1H-NMR. The unique up-field 1H-NMR chemical shift of TMSiPhe, highly efficient and specific incorporation of TMSiPhe enabled facile assignment of the TMSiPhe 1H-NMR signal, and characterization of multiple conformational state in a 150 kilodalton (kD) membrane protein complex, using only 5 M of protein and 20 min spectra accumulation time. This highly efficient and convenient methods should be broadly applicable for the investigation of dynamic conformation change of protein complexes.

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Liu, Q., He, Q.-t., Lyu, X.-x., Yang, F., Zhu, Z.-l., Xiao, P., Yang, Z., Zhang, F., Yang, Z.-y., Wang, X.-y., Sun, P., Wang, Q.-w., Qu, C.-x., Gong, Z., Lin, J.-Y., Xu, Z., Song, S.-l., Huang, S.-m., Guo, S.-c., Han, M.-j., Zhu, K.-k., Chen, X., Kahsai, A. W., Xiao, K.-H., Kong, W., Yu, X., Ruan, K., Li, F.-h., Niu, X.-g., Jin, C.-w., Wang, J., Sun, J.. 2019-12-19. Developing a DeSipher method for conformational change detection of membrane receptor complex at residue resolution. https://doi.org/10.1101/2019.12.18.873729

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