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Critical Role of a Loop at C-Terminal Domain on the Conformational Stability and Catalytic Efficiency of Chondroitinase ABC I

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Abstract

We used a combination of protein engineering and spectroscopic methods to investigate the effect of a long length loop on the conformational stability and activity of chondroitinase ABC I. This study involves manipulation of interactions around Asp689 as a key residue in the central region of the loop containing residues 681–695 located at C-terminal domain of the enzyme. According to the equilibrium unfolding experiments and considering thermodynamic m value and ΔG(H2O), we found that the folded state of H700N, L701T, and H700N/L701T are more compact relative to the folded state of wild-type protein and they become stabilized upon mutation. However, the compactness and stability of other variants are less than those of wild-type protein. According to enzyme activity measurements, we found that the catalytic efficiency of structurally stabilized variants is decreased, while that of destabilized mutants is improved.

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Acknowledgments

Financial support for this work was provided by research council of Tarbiat Modares University.

Conflict of interest

The authors have declared no conflict of interest.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    S. Akram Shirdel & Khosro Khajeh

  2. Department of Biology, Faculty of Sciences, University of Zanjan, Zanjan, Iran

    Khosrow Khalifeh

  3. Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Abolfazl Golestani

  4. Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Bijan Ranjbar

Authors
  1. S. Akram Shirdel
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  2. Khosrow Khalifeh
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  3. Abolfazl Golestani
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  4. Bijan Ranjbar
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  5. Khosro Khajeh
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Correspondence to Khosro Khajeh.

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Akram Shirdel, S., Khalifeh, K., Golestani, A. et al. Critical Role of a Loop at C-Terminal Domain on the Conformational Stability and Catalytic Efficiency of Chondroitinase ABC I. Mol Biotechnol 57, 727–734 (2015). https://doi.org/10.1007/s12033-015-9864-3

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