| Autor: |
H. W. Orton, I. D. Herath, A. Maleckis, S. Jabar, M. Szabo, B. Graham, C. Breen, L. Topping, S. J. Butler, G. Otting |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Magnetic Resonance, Vol 3, Pp 1-13 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2699-0016 |
| DOI: |
10.5194/mr-3-1-2022 |
| Popis: |
The metallo-β-lactamase IMP-1 features a flexible loop near the active site that assumes different conformations in single crystal structures, which may assist in substrate binding and enzymatic activity. To probe the position of this loop, we labelled the tryptophan residues of IMP-1 with 7-13C-indole and the protein with lanthanoid tags at three different sites. The magnetic susceptibility anisotropy (Δχ) tensors were determined by measuring pseudocontact shifts (PCSs) of backbone amide protons. The Δχ tensors were subsequently used to identify the atomic coordinates of the tryptophan side chains in the protein. The PCSs were sufficient to determine the location of Trp28, which is in the active site loop targeted by our experiments, with high accuracy. Its average atomic coordinates showed barely significant changes in response to the inhibitor captopril. It was found that localisation spaces could be defined with better accuracy by including only the PCSs of a single paramagnetic lanthanoid ion for each tag and tagging site. The effect was attributed to the shallow angle with which PCS isosurfaces tend to intersect if generated by tags and tagging sites that are identical except for the paramagnetic lanthanoid ion. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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