Abstract
We used xenon-perturbed 1H–15N multidimensional NMR to investigate the structural changes in the urea-induced equilibrium unfolding of the dimeric ketosteroid isomerase (KSI) from Pseudomonas putida biotype B. Three limited regions located on the β3-, β5- and β6-strands of dimeric interface were significantly perturbed by urea in the early stage of KSI unfolding, which could lead to dissociation of the dimer into structured monomers at higher denaturant concentration as the interactions in these regions are weakened. The results indicate that the use of xenon as an indirect probe for multidimensional NMR can be a useful method for the equilibrium unfolding study of protein at residue level.
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Acknowledgements
This work was supported by the grant from Korea Research Foundation (C00178). One of the authors (H.J.L) thanks Dr. Sung Jin Park and Dr. Woo Sung Son at National Research Laboratory of Membrane Protein Structure in Seoul National University for helpful discussion in backbone assignments.
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Lee, H.J., Moon, H.S., Jang, D.S. et al. Probing the equilibrium unfolding of ketosteroid isomerase through xenon-perturbed 1H–15N multidimensional NMR spectroscopy. J Biomol NMR 40, 65–70 (2008). https://doi.org/10.1007/s10858-007-9209-z
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DOI: https://doi.org/10.1007/s10858-007-9209-z