Red Edge Excitation Shift Spectroscopy is Highly Sensitive to Tryptophan Composition

William Kelton; Annmaree K. Warrender; Christopher Pudney; Jolyn Pan; Vickery Arcus

doi:10.1098/rsif.2023.0337

Red Edge Excitation Shift Spectroscopy is Highly Sensitive to Tryptophan Composition

William Kelton, Annmaree K. Warrender, Christopher Pudney, Jolyn Pan, Vickery Arcus

University of Waikato

Research output: Contribution to journal › Article › peer-review

3 Citations (SciVal)

Abstract

Red edge excitation shift (REES) spectroscopy relies on the unique emission profiles of fluorophore–solvent interactions to profile protein molecular dynamics. Recently, we reported the use of REES to compare the stability of 32 polymorphic IgG antibodies natively containing tryptophan reporter fluorophores. Here, we expand on this work to investigate the sensitivity of REES to variations in tryptophan content using a subset of IgG3 antibodies containing arginine to tryptophan polymorphisms. Structural analysis revealed that the additional tryptophan residues were situated in highly solvated environments. Subsequently, REES showed clear differences in fluorescence emission profiles when compared with the unmutated variants, thereby limiting direct comparison of their structural dynamics. These findings highlight the exquisite sensitivity of REES to minor variations in protein structure and tryptophan composition.

Original language	English
Article number	20230337
Number of pages	1
Journal	Journal of The Royal Society Interface
Volume	20
Issue number	208
Early online date	8 Nov 2023
DOIs	https://doi.org/10.1098/rsif.2023.0337
Publication status	Published - 8 Nov 2023

Funding

This work was supported by the Royal Society Te Apārangi (grant no.: 19-FRI-002); and the University of Waikato (Doctoral Scholarship).

Funders	Funder number
Royal Society Te Apārangi	19-FRI-002

Keywords

antibodies
protein dynamics
protein stability
red edge excitation shift
tryptophan fluorescence

ASJC Scopus subject areas

Bioengineering
Biophysics
Biochemistry
Biotechnology
Biomedical Engineering
Biomaterials

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10.1098/rsif.2023.0337Licence: CC BY

Cite this

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abstract = "Red edge excitation shift (REES) spectroscopy relies on the unique emission profiles of fluorophore–solvent interactions to profile protein molecular dynamics. Recently, we reported the use of REES to compare the stability of 32 polymorphic IgG antibodies natively containing tryptophan reporter fluorophores. Here, we expand on this work to investigate the sensitivity of REES to variations in tryptophan content using a subset of IgG3 antibodies containing arginine to tryptophan polymorphisms. Structural analysis revealed that the additional tryptophan residues were situated in highly solvated environments. Subsequently, REES showed clear differences in fluorescence emission profiles when compared with the unmutated variants, thereby limiting direct comparison of their structural dynamics. These findings highlight the exquisite sensitivity of REES to minor variations in protein structure and tryptophan composition.",

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AU - Kelton, William

AU - Warrender, Annmaree K.

AU - Pudney, Christopher

AU - Pan, Jolyn

AU - Arcus, Vickery

PY - 2023/11/8

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AB - Red edge excitation shift (REES) spectroscopy relies on the unique emission profiles of fluorophore–solvent interactions to profile protein molecular dynamics. Recently, we reported the use of REES to compare the stability of 32 polymorphic IgG antibodies natively containing tryptophan reporter fluorophores. Here, we expand on this work to investigate the sensitivity of REES to variations in tryptophan content using a subset of IgG3 antibodies containing arginine to tryptophan polymorphisms. Structural analysis revealed that the additional tryptophan residues were situated in highly solvated environments. Subsequently, REES showed clear differences in fluorescence emission profiles when compared with the unmutated variants, thereby limiting direct comparison of their structural dynamics. These findings highlight the exquisite sensitivity of REES to minor variations in protein structure and tryptophan composition.

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Red Edge Excitation Shift Spectroscopy is Highly Sensitive to Tryptophan Composition

Abstract

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Keywords

ASJC Scopus subject areas

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