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Unfolding and Refolding of Disulfide Proteins Using the Method Disulfide Scrambling

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Folding of Disulfide Proteins

Part of the book series: Protein Reviews ((PRON,volume 14))

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Abstract

Unfolding and refolding of disulfide proteins can be investigated by the method of disulfide scrambling which is based on the reversible conversion between the native (N) and scrambled isomers (X). The method of disulfide scrambling presents a number of unique features in elucidation of pathways of protein unfolding and refolding. (a) It allows trapping and isolation of diverse intermediates (unfolding and folding) for further structural analysis. (b) It demonstrates that protein denaturation and unfolding can be quantified independently. Denaturation is calculated by the conversion of native (N) to non-native X-isomers. Unfolding is measured by the progressive unfolding of X-isomers. (c) It shows that folding experiment can be initiated with a structurally defined X-isomer possessing the highest free energy among all unfolded X-isomers. (d) It reveals that the energy landscape of conformational heterogeneity (unfolding and refolding) can be illustrated by a diamond-shaped model. At two extreme ends of the energy landscape, the conformational heterogeneity is reduced to minimum.

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Abbreviations

α-LA:

α-Lactalbumin

HPLC:

High-pressure liquid chromatography

BPTI:

Bovine pancreatic trypsin inhibitor

TAP:

Tick anticoagulant peptide

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Authors and Affiliations

  1. Research Center for Protein Chemistry, Brown Foundation Institute of Molecular Medicine, 1825 Pressler Street, Houston, TX, 77030, USA

    Rowen J. Y. Chang

  2. Department of Biochemistry and Molecular Biology, The University of Texas, Houston, TX, 77030, USA

    Rowen J. Y. Chang

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  1. Rowen J. Y. Chang
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Correspondence to Rowen J. Y. Chang .

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Editors and Affiliations

  1. Brown Foundation Inst. of Molecular Medi, The University of Texas, Houston, 77030, Texas, USA

    Rowen J. Y. Chang

  2. Institut de Biotecnologia i de Biomed., Dept. de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, 08193, Spain

    Salvador Ventura

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Chang, R.J.Y. (2011). Unfolding and Refolding of Disulfide Proteins Using the Method Disulfide Scrambling. In: Chang, R., Ventura, S. (eds) Folding of Disulfide Proteins. Protein Reviews, vol 14. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-7273-6_5

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