A Genetically Encoded Trimethylsilyl 1D 1H-NMR Probe for Conformation Change in Large Membrane Protein Complexes
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.
Show 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.
Matching journals
The top 6 journals account for 50% of the predicted probability mass.
Similar papers in this journal
Similar papers in this journal
Similar papers in this journal
Similar papers in this journal
- A large-scale bioinformatic study of graspimiditides and structural characterization of albusimiditide 94%
- Integrative x-ray structure and molecular modeling for the rationalization of procaspase-8 inhibitor potency and selectivity 94%
- Uncovering the association mechanism between two intrinsically flexible proteins 94%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.