Abstract
Stopped-flow Fourier-transform infrared spectroscopy (SF-FTIR) was used to identify native as well as non-native secondary structures during the refolding of the calcium-binding protein α-lactalbumin. Infrared absorbance spectra were recorded in real time after a pH jump induced refolding of the protein. In the presence of calcium, the refolding is fast with concerted appearance of secondary structures; in its absence, folding is much slower and intricate, with transient formation and disappearance of non-native β-sheet. The possibility of detecting native as well as non-native structures at the same time is especially valuable in providing insight into the complexity of the refolding process of a protein.
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Acknowledgements
We thank F. Guillain and A. Sanson for careful reading of the manuscript, S. Crouzy for helping with Fig. 1, and E. Padros for the use of IR analysis software. A.T. was supported in part by a fellowship from Bio-Logic Co.
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Troullier, A., Reinstädler, D., Dupont, Y. et al. Transient non-native secondary structures during the refolding of α-lactalbumin detected by infrared spectroscopy. Nat Struct Mol Biol 7, 78–86 (2000). https://doi.org/10.1038/71286
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DOI: https://doi.org/10.1038/71286
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