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Solvent-Dependent Emissions Properties of a Model Aurone Enable Use in Biological Applications

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Abstract

Fluorophores are powerful visualization tools and the development of novel small organic fluorophores are in great demand. Small organic fluorophores have been derived from the aurone skeleton, 2-benzylidenebenzofuran-3(2H)-one. In this study, we have utilized a model aurone derivative with a methoxy group at the 3’ position and a hydroxyl group at the 4’ position, termed vanillin aurone, to develop a foundational understanding of structural factors impacting aurone fluorescence properties. The fluorescent behaviors of the model aurone were characterized in solvent environments differing in relative polarity and dielectric constant. These data suggested that hydrogen bonding or electrostatic interactions between excited state aurone and solvent directly impact emissions properties such as peak emission wavelength, emission intensity, and Stokes shift. Time-dependent Density Functional Theory (TD-DFT) model calculations suggest that quenched aurone emissions observed in water are a consequence of stabilization of a twisted excited state conformation that disrupts conjugation. In contrast, the calculations indicate that low polarity solvents such as toluene or acetone stabilize a brightly fluorescent planar state. Based on this, additional experiments were performed to demonstrate use as a turn-on probe in an aqueous environment in response to conditions leading to planar excited state stabilization. Vanillin aurone was observed to bind to a model ATP binding protein, YME1L, leading to enhanced emissions intensities with a dissociation equilibrium constant equal to ~ 30 µM. Separately, the aurone was observed to be cell permeable with significant toxicity at doses exceeding 6.25 µM. Taken together, these results suggest that aurones may be broadly useful as turn-on probes in aqueous environments that promote either a change in relative solvent polarity or through direct stabilization of a planar excited state through macromolecular binding.

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Accession Codes

The YME1L construct used in this study was derived from human YME1L (Uniprot Accession Code: Q96TA2 or National Center for Biotechnology Information Genbank Accession Code: CAB51858.1).

Data Availability

All datasets and materials relevant to the current study will be made available by the authors on request. Please contact the corresponding author with questions related to access of all relevant data and materials.

Abbreviations

ATP:

Adenosine triphosphate

AAA+:

ATPases Associated with various cellular Activities

YME1:

Yeast Mitochondrial Escape Protein 1

NLLS:

Nonlinear Least Squares

MD:

Molecular Dynamics

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Funding

This work was supported in part by funding to JMM/STH from the Middle Tennessee State University (MTSU) Department of Chemistry and the MTSU Molecular Biosciences (MOBI) Doctoral program. BA received financial support from the MTSU Department of Chemistry M.S. Degree program. DB received financial support from the MTSU Molecular Biosciences Doctoral program. This work used the Extreme Science and Engineering Discovery Environment (XSEDE) SDSC Expanse through allocation CHE180027.

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Authors and Affiliations

  1. Department of Chemistry, Middle Tennessee State University, 1301 East Main Street, Murfreesboro, TN, 37132, USA

    Beth Anderson, Chad Brambley, Scott T. Handy & Justin M. Miller

  2. Department of Biology, Middle Tennessee State University, 1301 East Main Street, Murfreesboro, TN, 37132, USA

    Daniel L. Bryant & Anthony Farone

  3. Department of Chemistry, Georgia State University, 50 Decatur Street SE, Atlanta, GA, 30302, USA

    Samer Gozem

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  1. Beth Anderson
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  2. Daniel L. Bryant
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Contributions

B.A., D.B., and J.M. wrote the first-draft of the manuscript and assisted in preparation of figures. B.A., D.B, S.G., and C.G. collected and analyzed experimental data for the manuscript. B.A., D.B., S.G., S.H., A.F., and J.M. collaboratively participated in the design of the study. All authors reviewed the manuscript.

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Correspondence to Justin M. Miller.

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Anderson, B., Bryant, D.L., Gozem, S. et al. Solvent-Dependent Emissions Properties of a Model Aurone Enable Use in Biological Applications. J Fluoresc (2024). https://doi.org/10.1007/s10895-024-03607-x

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