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Folding of cytochrome c initiated by submillisecond mixing

Nature Structural Biology volume 4, pages 44–50 (1997)Cite this article

Abstract

Cytochrome c folding was initiated using a new solution mixer that provides a time window which covers over 90% of the burst phase unresolved by conventional stop-flow measurements. Folding was followed by resonance Raman scattering. Kinetic analysis of the high frequency Raman data indicates that a nascent phase occurs within the mixing dead time of 100 μs. A significant fraction of the protein was found to be trapped in a misfolded bis-histidine form during the nascent phase at pH 4.5, thereby preventing the protein from folding rapidly and homogeneously. The nascent phase was followed by a haem-ligand exchange phase that populates the native histidine-methionine coordinated form through a thermodynamically controlled equilibrium.

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Author notes

  1. Satoshi Takahashi

    Present address: Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Kyoto, 606-01, Japan

Authors and Affiliations

  1. The Institute of Physical and Chemical Research (RIKEN), Wako, Saitama, 351-0, Japan

    Satoshi Takahashi

  2. Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, New York, 10461, USA

    Syun-Ru Yeh, Tapan K. Das, David S. Gottfried & Denis L. Rousseau

  3. Laboratory of Chemical Physics, Building 5, National Institutes of Health, Bethesda, Maryland, 20892-0520, USA

    Chi-Kin Chan

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  5. David S. Gottfried
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  6. Denis L. Rousseau
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Takahashi, S., Yeh, SR., Das, T. et al. Folding of cytochrome c initiated by submillisecond mixing. Nat Struct Mol Biol 4, 44–50 (1997). https://doi.org/10.1038/nsb0197-44

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