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Sensing Tryptophan Microenvironment of Amyloid Protein Utilizing Wavelength-Selective Fluorescence Approach

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Abstract

Structural transition among various forms of proteins involves subtle interplay between structure and dynamics and is crucial in human diseases. Red edge excitation shift (REES) represents a suitable approach to explore the environmental organization and dynamics surrounding tryptophan residues in proteins. Although REES from tryptophan residues has been reported for native, molten globule and denatured states of proteins, such data on the amyloid form of proteins is lacking. κ-casein is one of the most important constituents of casein micelles in milk and has a tendency to form amyloid fibril. We report here REES of the sole tryptophan residue for native, acid-denatured and urea-denatured forms of κ-casein. More importantly, we show that the amyloid form of κ-casein displays REES of 4 nm. We analyze these results in terms of tryptophan microenvironment in various forms of κ-casein, particularly the amyloid form. We conclude that REES is a sensitive tool to monitor structural plasticity in proteins.

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Acknowledgments

This work was supported by research grants from the Council of Scientific and Industrial Research, Govt. of India. A.C. gratefully acknowledges support from J.C. Bose Fellowship (Department of Science and Technology, Govt. of India). H.C. thanks the Council of Scientific and Industrial Research for the award of a Senior Research Associateship and the University Grants Commission for UGC-Assistant Professor position and UGC-Start-Up Grant (F.4-5(138-FRP)/2014(BSR)). A.C. is an Adjunct Professor of Tata Institute of Fundamental Research (Mumbai), RMIT University (Melbourne, Australia), Indian Institute of Technology (Kanpur), and Indian Institute of Science Education and Research (Mohali). We thank A. Harikrishna for help with transmission electron microscopy, G. Aditya Kumar for help in making figures, and members of the Chattopadhyay laboratory for their comments and discussions.

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Authors and Affiliations

  1. CSIR-Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007, India

    Hirak Chakraborty & Amitabha Chattopadhyay

  2. School of Chemistry, Sambalpur University, Jyoti Vihar, Burla, Odisha, 768 019, India

    Hirak Chakraborty

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  1. Hirak Chakraborty
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  2. Amitabha Chattopadhyay
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Correspondence to Amitabha Chattopadhyay.

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Chakraborty, H., Chattopadhyay, A. Sensing Tryptophan Microenvironment of Amyloid Protein Utilizing Wavelength-Selective Fluorescence Approach. J Fluoresc 27, 1995–2000 (2017). https://doi.org/10.1007/s10895-017-2138-7

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  • DOI: https://doi.org/10.1007/s10895-017-2138-7

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