Abstract
ModA is a soluble periplasmic molybdate-binding protein found in most gram-negative bacteria. It is part of the ABC transporter complex ModABC that moves molybdenum into the cytoplasm, to be used by enzymes that carry out various redox reactions. Since there is no clear analog for ModA in humans, this protein could be a good target for antibacterial drug design. Backbone 1H, 13C and 15N chemical shifts of apo and molybdate-bound ModA from E. coli were assigned at pHs 6.0 and 4.5. In addition, side chain atoms were assigned for apo ModA at pH 6.0. When comparing apo and molybdate-bound ModA at pH 6.0, large chemical shift perturbations are observed, not only in areas near the bound metal, but also in regions that are distant from the metal-binding site. Given the significant conformational change between apo and holo ModA, we might expect the large chemical shift changes to be more widespread; however, since they are limited to specific regions, the residues with large perturbations may reveal allosteric sites that could ultimately be important for the design of antibiotics that target ModA.
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Data availability
Chemical shift assignment data have been deposited to the BioMagResBank (BMRB) with accession numbers 52193 and 52200.
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Acknowledgements
Thanks go to Nicholas Mead for assistance with protein expression and purification. We are grateful to the National Institute of General Medical Sciences of the National Institutes of Health (award number SC3GM116745) for supporting this research, and for a Major Research Instrumentation grant to purchase a 600 MHz NMR spectrometer provided by the National Science Foundation (award number CHE-1040134).
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H.N. made the protein samples, performed the majority of the chemical shift assignments, wrote portions of the manuscript and made part of Fig. 2; K.C. performed some of the assignments, deposited the data to the BMRB, wrote the majority of the manuscript and made the remaining figures. Both authors reviewed the manuscript.
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Nguyen, H.L., Crowhurst, K.A. Solution NMR chemical shift assignment of apo and molybdate-bound ModA at two pHs. Biomol NMR Assign 18, 93–98 (2024). https://doi.org/10.1007/s12104-024-10173-7
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DOI: https://doi.org/10.1007/s12104-024-10173-7