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Probing the interaction between human serum albumin and the sodium dodecyl sulphate with fluorescence correlation spectroscopy

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Abstract

The denaturation of human serum albumin (HSA) upon interaction with the surfactant sodium dodecyl sulphate (SDS) was examined by measuring the diffusion time of fluorophore (RITC) tagged HSA under near single-molecule conditions using fluorescence correlation spectroscopy. The diffusion time shows four distinct regions as a function of SDS concentration, which corresponds to (I) opening of the tertiary structure, (II) non-specific SDS aggregation, (III) opening of the secondary structure, and (IV) aggregation of SDS around the secondary structure. Diffusion time increases from 383 µs for the free protein to 1002 µs for the SDS bound protein, which leads to an effective increase in the hydrodynamic radius by a factor of about 2.6.

Graphic abstract

Fluorescence correlation spectroscopy reveals a four-step interaction regime between SDS and HSA. The initial opening of the tertiary structure, followed by non-specific aggregation and opening up the secondary structure and finally leading to formation of protein bound micelles with an effective increase of hydrodynamic radius by a factor of 2.6.

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Acknowledgements

Authors wish to thank Prof. Sudipta Maiti for his help in setting up the FCS instrument as part of the first FCS workshop held at TIFR in the year 2009. The FCS instrument was procured as part of the Department of Information Technology Govt. of India sponsored research project entitled Construction and multi-site commissioning of multiple fluorescence correlation spectrometers [Project No. 12(4)/2007-PDD]. VS gratefully acknowledges CSIR for fellowship.

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  1. Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai, Maharashtra, 400 076, India

    Vaishali Samant, Arghya Dey & G Naresh Patwari

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  1. Vaishali Samant
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  2. Arghya Dey
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  3. G Naresh Patwari
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Correspondence to Vaishali Samant or G Naresh Patwari.

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Samant, V., Dey, A. & Naresh Patwari, G. Probing the interaction between human serum albumin and the sodium dodecyl sulphate with fluorescence correlation spectroscopy. J Chem Sci 132, 109 (2020). https://doi.org/10.1007/s12039-020-01816-y

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  • DOI: https://doi.org/10.1007/s12039-020-01816-y

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