Abstract
The aminoglycoside phosphotransferase(3′)-IIIa (APH) is a promiscuous enzyme and renders a large number of structurally diverse aminoglycoside antibiotics useless against infectious bacteria. A remarkable property of this ~31 kDa enzyme is in its unusual dynamic behavior in solution; the apo-form of the enzyme exchanges all of its backbone amide protons within 15 h of exposure to D 2 O while aminoglycoside-bound forms retain ~40% of the amide protons even after >90 h of exposure. Moreover, the number of observable peaks and their dispersion in HSQC spectra varies with each aminoglycoside, rendering the resonance assignments very challenging. Therefore, the binary APH–tobramycin complex, which shows the largest number of well-resolved peaks, was used for the backbone resonance assignments (Cα, C, N, H, and some Cβ) of this protein (BMRB-16337).
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Acknowledgments
This research was supported by a Grant from the National Science Foundation (mostly through MCB 01110741 and a small extend through MCB-0842743 to EHS), the Center of Excellence for Structural Biology, and the Hunsicker Research Award through the Department of Biochemistry Cellular and Molecular Biology at the University of Tennessee. This research benefited from activities at the Southeast Collaboratory for High-Field Biomolecular NMR, a research resource at the University of Georgia, funded by the National Institute of General Medical Sciences (NIGMS grant number GM66340) and the Georgia Research Alliance.
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Serpersu, E.H., Özen, C., Norris, A.L. et al. Backbone resonance assignments of a promiscuous aminoglycoside antibiotic resistance enzyme; the aminoglycoside phosphotransferase(3′)-IIIa. Biomol NMR Assign 4, 9–12 (2010). https://doi.org/10.1007/s12104-009-9195-z
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DOI: https://doi.org/10.1007/s12104-009-9195-z