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Structural Aspects of a Protein–Surfactant Assembly: Native and Reduced States of Human Serum Albumin

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Abstract

The inherently present seventeen disulfide bonds of the circulatory protein, human serum albumin (HSA) provide the necessary structural stability. Various spectroscopic approaches were used to investigate the effect of reduction of these disulfide bonds and its binding with the anionic surfactant, sodium dodecyl sulfate (SDS). Based on several spectroscopic analyses, our investigations highlight the following interesting aspects: (1) HSA on reduction loses not only its tertiary structure but also a significant amount of secondary structure as well. However, the reduced state of the protein is not like the molten-globule, (2) this structural loss of the protein due to reduction is more prominent than that caused by higher SDS concentrations alone and can certainly be attributed to the role of disulfide bonds, (3) lower surfactant concentrations provide marginal structural rigidity to the native state of the protein, whereas, higher concentrations of SDS induces secondary structure to the reduced state of HSA, (4) the binding of SDS with both the native and reduced states of HSA, occurred in three distinct stages which was followed by a saturation stage. However, the nature of such binding is different for both the states as investigated by using the Stern–Volmer equations and estimating the thermodynamic parameters. Besides, in contrast to the native state, the reduced state of HSA shows that the lone tryptophan residue gets more buried. However, there occurs a sudden decrement in the lifetime of the tryptophan and the hydrodynamic diameter increases by twofold.

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Abbreviations

HSA:

Human serum albumin

SDS:

Sodium dodecyl sulfate

BSA:

Bovine serum albumin

DTT:

Dithiothretol

Trp:

Tryptophan

Cys:

Cysteine

DLS:

Dynamic light scattering

CD:

Circular dichroism

DTNB:

5,5′-Dithiobis(2-nitrobenzoic acid)

UV–Vis:

Ultra violet–visible

dH:

Hydrodynamic diameter

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Acknowledgments

We thank Professor Vinod Kumar Singh, Director IISER Bhopal for his constant encouragement and support. SM sincerely thanks Professor Sandeep Verma, IIT Kanpur for many stimulating discussions and suggestions. We sincerely thank Professor A. Dasgupta, University of Calcutta for allowing us to use the DLS instrumental facility. We are grateful to Professor Soumen Basak, Chemical Science Division, SINP, Kolkata for his guidance during the CD measurements. We also thank Dr. Nabendu S. Chatterjee, Biochemistry Division, National Institute of Cholera and Enteric Diseases, Kolkata, for FPLC (Fast Protein Liquid Chromatography) instrumental facility. UA thanks CSIR, SG thanks UGC for providing fellowship and SR thanks University of Calcutta for providing financial support. SM thanks the DST-Fast track scheme (No.: SR/FT/CS-19/2011) SERB for financial support.

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    Authors and Affiliations

    1. Department of Chemistry, Indian Institute of Science Education and Research Bhopal, ITI Campus (Gas Rahat) Building, Govindpura, Bhopal, 462 023, Madhya Pradesh, India

      Uttam Anand, Subhadip Ghosh & Saptarshi Mukherjee

    2. Department of Biotechnology and Dr. B. C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, 700 019, India

      Sutapa Ray & Rajat Banerjee

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    1. Uttam Anand
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    Correspondence to Rajat Banerjee or Saptarshi Mukherjee.

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    Uttam Anand and Sutapa Ray have contributed equally to this work.

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    Anand, U., Ray, S., Ghosh, S. et al. Structural Aspects of a Protein–Surfactant Assembly: Native and Reduced States of Human Serum Albumin. Protein J 34, 147–157 (2015). https://doi.org/10.1007/s10930-015-9606-1

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