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Heat, pH Induced Aggregation and Surface Hydrophobicity of S. cerevesiae Ssa1 Protein

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Abstract

Heat shock protein 70 is a conserved protein among organisms. Hsp70 helps substrate proteins to fold correctly. Unfolded substrate proteins increase the probability of the aggregate formation. High level recombinant protein expression in biotechnology often leads insoluble inclusion bodies. To prevent aggregation and to obtain high levels of soluble proteins, Hsp co-expression with desired recombinant protein in yeast becomes a popular method. For this purpose, S. cerevesiae cytosolic Hsp70 (Ssa1) biochemical properties were characterized. Alteration of Ssa1 structure between ATP- and ADP-bound states regulates its function. Therefore, conformation-dependent Ssa1 hydrophobicity and as a result aggregation may also play a key role in Ssa1 function. Therefore, a combination of FTIR, acrylamide quenching, and ANS was used to investigate the effect of nucleotide binding on the structure of Ssa1. Ssa1 secondary structure alterations and hydrophobic properties in aqueous solutions with differing ionic strengths and temperature were also studied.

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Abbreviations

Hsp:

Heat shock protein

FTIR:

Fourier transform infrared spectroscopy

ANS:

8-Anilino-2-naphthyl sulfonic acid

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Acknowledgments

This work was funded partly by the Turkish Planning Organization (Grant DPT-K.120220-2006) and through seed grants from the Turkish National Academy of Sciences (TUBA-GEBIP) and from Cumhuriyet University Graduate School for Derya Arslan.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Medicine, Cumhuriyet University, 58140, Sivas, Turkey

    Yusuf Tutar

  2. Department of Chemistry, Faculty of Science, Cumhuriyet University, 58140, Sivas, Turkey

    Derya Arslan

  3. Department of Biology, Faculty of Science, Ankara University, 06100, Ankara, Turkey

    Lütfi Tutar

Authors
  1. Yusuf Tutar
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  2. Derya Arslan
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  3. Lütfi Tutar
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Correspondence to Yusuf Tutar.

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Tutar, Y., Arslan, D. & Tutar, L. Heat, pH Induced Aggregation and Surface Hydrophobicity of S. cerevesiae Ssa1 Protein. Protein J 29, 501–508 (2010). https://doi.org/10.1007/s10930-010-9280-2

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