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Chaperones rescue luciferase folding by separating its domains.

Publication ,  Journal Article
Scholl, ZN; Yang, W; Marszalek, PE
Published in: The Journal of biological chemistry
October 2014

Over the last 50 years, significant progress has been made toward understanding how small single-domain proteins fold. However, very little is known about folding mechanisms of medium and large multidomain proteins that predominate the proteomes of all forms of life. Large proteins frequently fold cotranslationally and/or require chaperones. Firefly (Photinus pyralis) luciferase (Luciferase, 550 residues) has been a model of a cotranslationally folding protein whose extremely slow refolding (approximately days) is catalyzed by chaperones. However, the mechanism by which Luciferase misfolds and how chaperones assist Luciferase refolding remains unknown. Here we combine single-molecule force spectroscopy (atomic force microscopy (AFM)/single-molecule force spectroscopy) with steered molecular dynamic computer simulations to unravel the mechanism of chaperone-assisted Luciferase refolding. Our AFM and steered molecular dynamic results show that partially unfolded Luciferase, with the N-terminal domain remaining folded, can refold robustly without chaperones. Complete unfolding causes Luciferase to get trapped in very stable non-native configurations involving interactions between N- and C-terminal residues. However, chaperones allow the completely unfolded Luciferase to refold quickly in AFM experiments, strongly suggesting that chaperones are able to sequester non-natively contacting residues. More generally, we suggest that many chaperones, rather than actively promoting the folding, mimic the ribosomal exit tunnel and physically separate protein domains, allowing them to fold in a cotranslational-like sequential process.

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Published In

The Journal of biological chemistry

DOI

EISSN

1083-351X

ISSN

0021-9258

Publication Date

October 2014

Volume

289

Issue

41

Start / End Page

28607 / 28618

Related Subject Headings

  • Thermodynamics
  • Reticulocytes
  • Recombinant Fusion Proteins
  • Rabbits
  • Protein Unfolding
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Refolding
  • Protein Denaturation
  • Molecular Dynamics Simulation
 

Citation

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Scholl, Z. N., Yang, W., & Marszalek, P. E. (2014). Chaperones rescue luciferase folding by separating its domains. The Journal of Biological Chemistry, 289(41), 28607–28618. https://doi.org/10.1074/jbc.m114.582049
Scholl, Zackary N., Weitao Yang, and Piotr E. Marszalek. “Chaperones rescue luciferase folding by separating its domains.The Journal of Biological Chemistry 289, no. 41 (October 2014): 28607–18. https://doi.org/10.1074/jbc.m114.582049.
Scholl ZN, Yang W, Marszalek PE. Chaperones rescue luciferase folding by separating its domains. The Journal of biological chemistry. 2014 Oct;289(41):28607–18.
Scholl, Zackary N., et al. “Chaperones rescue luciferase folding by separating its domains.The Journal of Biological Chemistry, vol. 289, no. 41, Oct. 2014, pp. 28607–18. Epmc, doi:10.1074/jbc.m114.582049.
Scholl ZN, Yang W, Marszalek PE. Chaperones rescue luciferase folding by separating its domains. The Journal of biological chemistry. 2014 Oct;289(41):28607–28618.

Published In

The Journal of biological chemistry

DOI

EISSN

1083-351X

ISSN

0021-9258

Publication Date

October 2014

Volume

289

Issue

41

Start / End Page

28607 / 28618

Related Subject Headings

  • Thermodynamics
  • Reticulocytes
  • Recombinant Fusion Proteins
  • Rabbits
  • Protein Unfolding
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Refolding
  • Protein Denaturation
  • Molecular Dynamics Simulation