Abstract
Trichobakin (TBK) is a type-I ribosome-inactivating protein (RIP-I), acting as an extremely potent inhibitor of protein synthesis in the cell-free translation system of rabbit reticulocyte lysate (IC50: 3.5 pM). In this respect, TBK surpasses the well-studied highly homologous RIP-I trichosanthin (IC50: 20–27 pM), therefore creation of recombinant toxins based on it is of great interest. TBK needs to penetrate into cytosol through the cell membrane and specifically bind to α-sarcin/ricin loop of 28S ribosome RNA to perform the function of specific RNA depurination. At the moment, there is no detailed structural-dynamic information in solution about diverse states RIP-I can adopt at different stages on the way to protein synthesis inhibition. In this work, we report a near-complete assignment of 1H, 13C, and 15N TBK (27.3 kDa) resonances and analysis of the secondary structure based on the experimental chemical shifts data. This work will serve as a basis for further investigations of the structure, dynamics and interactions of the TBK with its molecular partners using NMR techniques.
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Acknowledgements
These studies were supported by the Russian Science Foundation (project 19-74-30014) in part of NMR spectroscopy measurements, by Belarusian Republican Foundation for Fundamental Research (project BRFBR-VASTX17B-012) and by the Vietnam Academy of Science and Technology (project VAST.HTQT.BELARUS.02/17-18) in part of protein preparation and resonance assignments.
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Britikov, V.V., Britikova, E.V., Urban, A.S. et al. Backbone and side-chain chemical shift assignments for the ribosome-inactivating protein trichobakin (TBK). Biomol NMR Assign 14, 55–61 (2020). https://doi.org/10.1007/s12104-019-09920-y
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DOI: https://doi.org/10.1007/s12104-019-09920-y