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The Effects of Molecular Crowding on the Amyloid Fibril Formation of α-Lactalbumin and the Chaperone Action of α-Casein

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Abstract

Amyloid fibrils arise from the slow aggregation of intermediately folded protein states. In this study the kinetics of the protein fibril formation of α-lactalbumin and its prevention by αS-casein in the presence and absence of the crowding agent, dextran (68 kDa), have been compared using a thioflavin T binding assay. It was found that αS-casein, a molecular chaperone found in bovine milk, is a potent in vitro inhibitor of α-lactalbumin fibrillization. The effect of αS-casein in preventing fibril formation was significant, although less than it is in the absence of the crowding agent, dextran. The interaction between the chaperone and the α-lactalbumin and structural change in the target protein are also shown using intrinsic fluorescence intensity, an ANS binding assay, CD spectroscopy and size-exclusion HPLC. In summary, α-casein interacts with α-lactalbumin and prevents amyloid formation but not as well as it does when the crowding agent, dextran, not present.

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Abbreviations

ThT:

Thioflavin T

ANS:

1-anilino-8-naphthalene sulfonic acid

CD:

Circular dichroism

DTT:

Deuterated dithiothreitol

HPLC:

High performance liquid chromatography

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, University of Sistan and Baluchestan, Zahedan, Iran

    Arezou Ghahghaei & Nasim Faridi

  2. Department of Biological Sciences, Tarbiat Moallem University, Tehran, Iran

    Adeleh Divsalar

Authors
  1. Arezou Ghahghaei
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  2. Adeleh Divsalar
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  3. Nasim Faridi
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Correspondence to Arezou Ghahghaei.

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Ghahghaei, A., Divsalar, A. & Faridi, N. The Effects of Molecular Crowding on the Amyloid Fibril Formation of α-Lactalbumin and the Chaperone Action of α-Casein. Protein J 29, 257–264 (2010). https://doi.org/10.1007/s10930-010-9247-3

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  • DOI: https://doi.org/10.1007/s10930-010-9247-3

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