Abstract
The E.coli maltose binding protein (MBP) is a 42.5 kDa molecule widely employed in many biotechnology applications. Because of its molecular size, it has become the main model system for the development of solution NMR methods adapted to large biomolecular targets. Here, we report virtually complete (~ 90%) backbone resonance assignments obtained on a microcrystalline sample of MBP with 1H-detected solid-state NMR at fast (> 100 kHz) magic-angle spinning. We additionally present the detailed description of the methodology employed for the preparation of the sample and the acquisition and analysis of the NMR spectra. The chemical shifts, obtained with a single uniformly 15N, 13C-labelled and fully-protonated sample and about 2 weeks on a 800 MHz NMR spectrometer, have been deposited to the BMRB under the accession number 50089.
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Acknowledgements
The expression vector was a gift by Prof. Gottfried Otting (ANU). The work was funded by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (ERC-2015-CoG GA 648974). J.S. was supported by the European Commission’s REA with a MSCA fellowship (GA 661799).
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1H, 13C and 15N backbone and side-chain chemical shifts of crystalline MBP have been deposited in the BioMagResBank (https://www.bmrb.wisc.edu) under the accession number 50089.
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Schubeis, T., Stanek, J. & Pintacuda, G. Backbone assignment of crystalline E. coli maltose binding protein. Biomol NMR Assign 15, 317–322 (2021). https://doi.org/10.1007/s12104-021-10023-w
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DOI: https://doi.org/10.1007/s12104-021-10023-w