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A Partially Folded State of Ovalbumin at Low pH Tends to Aggregate

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Abstract

At pH 2, ovalbumin retains native-like secondary structure as seen by far-UV CD and FTIR, but lacks well-defined tertiary structure as seen by the fluorescence and near-UV CD spectra. Addition of 20 mM Trifluoroacetic acid (TFA) or 30 mM Trichloroacetic acid (TCA) on acid-induced state results in protein aggregation. This aggregated state possesses extensive β-sheet structure as revealed by far-UV CD and FTIR spectroscopy. Furthermore, the aggregates exhibit decreased ANS fluorescence and increased thioflavin T fluorescence. The presence of aggregates was confirmed by size exclusion chromatography. Such a formation of β-sheet structure is found in the amyloid of a number of neurological diseases such as Alzheimer’s and scrapie. Ovalbumin at low pH, in the presence of K2SO4, exists in partially folded state characterized by native-like secondary structure and tertiary folds.

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Abbreviations

ANS:

8-Anilino-1-naphthalene-sulfonic acid

CD:

Circular dichroism

Gdn-HCl:

Guanidine hydrochloride

ATR-FTIR:

Attenuated total reflection Fourier transform infrared spectroscopy

K2SO4 :

Potassuim sulfate

MRE:

Mean residual ellipticity

SEC:

Size exclusion chromatography

TFA:

Trifluoroacetic acid

TCA:

Trichloroacetic acid

Th T:

Thioflavin T

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Acknowledgments

The authors are highly thankful for the facilities obtained at AMU Aligarh. Financial support from the Department of Science and Technology, New Delhi in the form of project (SR/LS-087/2007) and CSIR in the form of project No. 37(1365)/09/EMR-II is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Life Science, AMU, Aligarh, 202 002, India

    Aabgeena Naeem, Taqi Ahmed Khan, Mohammad Muzaffar & Saman Ahmad

  2. Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, 202 002, India

    M. Saleemuddin

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  1. Aabgeena Naeem
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  2. Taqi Ahmed Khan
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Correspondence to Aabgeena Naeem.

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Naeem, A., Khan, T.A., Muzaffar, M. et al. A Partially Folded State of Ovalbumin at Low pH Tends to Aggregate. Cell Biochem Biophys 59, 29–38 (2011). https://doi.org/10.1007/s12013-010-9108-x

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