Abstract
The internal dynamics of recombinant Major Urinary Protein (rMUP) have been investigated by monitoring transverse nitrogen-15 relaxation using multiple-echo Carr–Purcell–Meiboom–Gill (CPMG) experiments. While the ligand-free protein (APO-rMUP) features extensive evidence of motions on the milliseconds time scale, the complex with 2-methoxy-3-isobutylpyrazine (HOLO-rMUP) appears to be much less mobile on this time scale. At 308 K, exchange rates k ex = 500–2000 s−1 were typically observed in APO-rMUP for residues located adjacent to a β-turn comprising residues 83–87. These residues occlude an entry to the binding pocket and have been proposed to be a portal for ligand entry in other members of the lipocalin family, such as the retinol binding protein and the human fatty-acid binding protein. Exchange rates and populations are largely uncorrelated, suggesting local ‘breathing’ motions rather than a concerted global conformational change.
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Acknowledgements
We are indebted to Dr Julien Wist (Bogotá, Columbia) for stimulating discussions. This work was supported by the Swiss National Science Foundation, the Swiss Commission for Innovation and Technology, the French CNRS, and the BBSRC (grant no. B19388 to SWH, United Kingdom).
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Perazzolo, C., Verde, M., Homans, S.W. et al. Evidence of chemical exchange in recombinant Major Urinary Protein and quenching thereof upon pheromone binding. J Biomol NMR 38, 3–9 (2007). https://doi.org/10.1007/s10858-006-9110-1
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DOI: https://doi.org/10.1007/s10858-006-9110-1