2D-IR Spectroscopy of Proteins in H2O - A Perspective

Rutherford, Samantha H, Baker, Matthew orcid iconORCID: 0000-0003-2362-8581 and Hunt, Neil T (2023) 2D-IR Spectroscopy of Proteins in H2O - A Perspective. The Journal of Chemical Physics . ISSN 0021-9606

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Official URL: https://doi.org/10.1063/5.0129480


The form of the amide I infrared absorption band provides a sensitive probe of the secondary structure and dynamics of proteins in the solution phase. However, the frequency coincidence of the amide I band with the bending vibrational mode of H­2O has necessitated the widespread use of deuterated solvents. Recently, it has been demonstrated that ultrafast 2D-IR spectroscopy allows the detection of the protein amide I band in H2O-based fluids, meaning that IR methods can now be applied to study proteins in physiologically relevant solvents. In this perspective, we describe the basis of the 2D-IR method for observing the protein amide I band in H2O and show how this development has the potential to impact on areas ranging from our fundamental appreciation of protein structural dynamics to new applications for 2D-IR spectroscopy in the analytical and biomedical sciences. In addition, we discuss how the spectral response of water, rather than being a hindrance, now provides a basis for new approaches to data pre-processing, standardisation of 2D-IR data collection and signal quantification. Ultimately, we visualise a direction of travel towards the creation of 2D-IR spectral libraries that can be linked to advanced computational methods for use in high-throughput protein screening and disease diagnosis.

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