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Disrupting Fluorescence by Mutagenesis in a Green Fluorescent Fatty Acid Binding Protein from a Marine Eel

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Abstract

Biofluorescence has been found to be an increasingly widespread phenomenon in the ocean. The reclusive Caribbean chlopsid eel, Kaupichthys hyoproroides displays bright green fluorescence in its native marine environment. We have previously shown the fluorescence to be attributed to a fluorescent fatty acid-binding protein, Chlopsid FP, part of a larger family of fluorescent fatty acid-binding proteins, including the homologous UnaG. All require the addition of exogenous bilirubin for fluorescence. Here, we report the generation of a series of point mutants, and deletions that result in the quenching of fluorescence in Chlopsid FP. In addition, we report the binding constants of bilirubin to Chlopsid FP and mutants, measured by fluorescence titration. This study provides key insights into the potential mechanism of fluorescence in this class of fluorescent fatty acid-binding proteins.

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Acknowledgements

This work was supported by a National Science Foundation Career Grant to J.P.G (Award Number MCB 1652731). We would like to thank Professor Pablo Peixoto for use of his fluorimeter.

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Authors and Affiliations

  1. Department of Natural Sciences, Baruch College, City University of New York, New York, NY, 10010, USA

    Sara Rose Krivoshik, Andrew M. Guarnaccia, David F. Gruber & Jean P. Gaffney

  2. The Graduate Center, Program in Biology, City University of New York, New York, NY, 10016, USA

    Sara Rose Krivoshik, Andrew M. Guarnaccia, David F. Gruber & Jean P. Gaffney

  3. Department of Chemistry, Saint Peter’s University, Jersey City, NJ, 07306, USA

    Daniel B. Fried

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  1. Sara Rose Krivoshik
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  2. Andrew M. Guarnaccia
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  5. Jean P. Gaffney
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Correspondence to Jean P. Gaffney.

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Krivoshik, S.R., Guarnaccia, A.M., Fried, D.B. et al. Disrupting Fluorescence by Mutagenesis in a Green Fluorescent Fatty Acid Binding Protein from a Marine Eel. Protein J 39, 145–151 (2020). https://doi.org/10.1007/s10930-020-09883-3

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  • DOI: https://doi.org/10.1007/s10930-020-09883-3

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