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Site-Specific Interrogation of Protein Structure and Stability

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Protein Folding

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2376))

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Abstract

To execute their function or activity, proteins need to possess variability in local electrostatic environment, solvent accessibility, structure, and stability. However, assessing any protein property in a site-specific manner is not easy since native spectroscopic signals often lack the needed specificity. One strategy that overcomes this limitation is to use unnatural amino acids that exhibit distinct spectroscopic features. In this chapter, we describe several such unnatural amino acids (UAAs) and their respective applications in site-specific interrogation of protein structure and stability using standard biophysical methods, including circular dichroism (CD), infrared (IR), and fluorescence spectroscopies.

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Acknowledgments

We gratefully acknowledge financial support from the National Institutes of Health (GM-065978 and P41-GM104605). I.A.A. is supported by a NIH T32 Interdisciplinary Cardiovascular Training Grant (T32-HL007954).

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Author notes

  1. Debopreeti Mukherjee and Ismail A. Ahmed contributed equally with all other contributors.

Authors and Affiliations

  1. Department of Chemistry, University of Pennsylvania, Philadelphia, PA, USA

    Debopreeti Mukherjee & Feng Gai

  2. Department of Biochemistry and Molecular Biophysics, University of Pennsylvania, Philadelphia, PA, USA

    Ismail A. Ahmed

Authors
  1. Debopreeti Mukherjee
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  2. Ismail A. Ahmed
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  3. Feng Gai
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Correspondence to Feng Gai .

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Editors and Affiliations

  1. Department of Bioengineering and Center for Cellular and Biomolecular Machines, University of California, Merced, Merced, CA, USA

    Victor Muñoz

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Mukherjee, D., Ahmed, I.A., Gai, F. (2022). Site-Specific Interrogation of Protein Structure and Stability. In: Muñoz, V. (eds) Protein Folding. Methods in Molecular Biology, vol 2376. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1716-8_3

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  • DOI: https://doi.org/10.1007/978-1-0716-1716-8_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1715-1

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