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Co-translational domain folding as the structural basis for the rapid de novo folding of firefly luciferase

Nature Structural Biology volume 6, pages 697–705 (1999)Cite this article

Abstract

The 62 kDa protein firefly luciferase folds very rapidly upon translation on eukaryotic ribosomes. In contrast, the chaperone-mediated refolding of chemically denatured luciferase occurs with significantly slower kinetics. Here we investigate the structural basis for this difference in folding kinetics. We find that an N-terminal domain of luciferase (residues 1–190) folds co-translationally, followed by rapid formation of native protein upon release of the full-length polypeptide from the ribosome. In contrast sequential domain formation is not observed during in vitro refolding. Discrete unfolding steps, corresponding to domain unfolding, are however observed when the native protein is exposed to increasing concentrations of denaturant. Thus, the co-translational folding reaction bears more similarities to the unfolding reaction than to refolding from denaturant. We propose that co-translational domain formation avoids intramolecular misfolding and may be critical in the folding of multidomain proteins.

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Figure 1: A protease-resistant luciferase domain forms co-translationally.
Figure 2: Firefly luciferase unfolds in a domainwise manner upon chemical denaturation.
Figure 3: Identification of the co-translationally folded domain as the 22 kDa N-terminal fragment of luciferase.
Figure 4: The 22 kDa domain is not observed during the chaperone-mediated refolding of denatured luciferase.
Figure 5: Position of different domains in the luciferase crystal structure.

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Acknowledgements

J.F. is supported by an NIH grant. P.T. is supported by an NSF grant and an NCI grant to the Sloan-Kettering Structural Chemistry Laboratory.

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Authors and Affiliations

  1. Department of Biological Sciences, Stanford University, Stanford, 94305, California, USA

    Judith Frydman

  2. Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, 10021, New York, USA

    Hediye Erdjument-Bromage & Paul Tempst

  3. Department of Cellular Biochemistry, Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, Martinsried, D-82152, Germany

    F. Ulrich Hartl

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Correspondence to Judith Frydman.

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Frydman, J., Erdjument-Bromage, H., Tempst, P. et al. Co-translational domain folding as the structural basis for the rapid de novo folding of firefly luciferase. Nat Struct Mol Biol 6, 697–705 (1999). https://doi.org/10.1038/10754

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