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Evidence for an obligatory intermediate in the folding of lnterleukin-1β

Nature Structural Biology volume 4, pages 725731 (1997) | Download Citation

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Abstract

The folding of the β-sheet protein, interleukin-1β, was examined at pH 5.0 and 25 °C using pulse-labelling hydrogen exchange and electrospray ionization mass spectrometric analysis, as well as stopped-flow circular dichroism and fluorescence spectroscopies. The first detectable event is the formation of a partially folded intermediate in a kinetic step with a relaxation time of 126 ± 26 ms. There is a lag in native protein production of at least 400 ms. Optical studies indicate that the intermediate is converted to the native species in a reaction with a relaxation time of 43 ± 5 s. The kinetic rates determined from stopped-flow fluorescence, circular dichroism and pulse-labelling experiments are similar and consistent with a simple sequential model for the folding pathway of interleukin-1β at pH 5.0 and 25 °C. Taken together, our data provide kinetic evidence that formation of the native state of interleukin-1β proceeds through an obligatory intermediate. We explain our results in terms of the classical and new views of protein folding.

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  1. Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California 92093-0359, USA.

    • David K. Heidary
    • , Melinda Roy
    •  & Patricia A. Jennings
  2. Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093-0647, USA.

    • Larry A. Gross

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Correspondence to Patricia A. Jennings.

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DOI

https://doi.org/10.1038/nsb0997-725

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