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The structural basis for red fluorescence in the tetrameric GFP homolog DsRed

Nature Structural Biology volume 7, pages 1133–1138 (2000)Cite this article

Abstract

Green fluorescent protein (GFP) has rapidly become a standard tool for investigating a variety of cellular activities, and has served as a model system for understanding spectral tuning in chromophoric proteins. Distant homologs of GFP in reef coral and anemone display two new properties of the fluorescent protein family: dramatically red-shifted spectra, and oligomerization to form tetramers. We now report the 1.9 Å crystal structure of DsRed, a red fluorescent protein from Discosoma coral. DsRed monomers show similar topology to GFP, but additional chemical modification to the chromophore extends the conjugated π-system and likely accounts for the red-shifted spectra. Oligomerization of DsRed occurs at two chemically distinct protein interfaces to assemble the tetramer. The DsRed structure reveals the chemical basis for the functional properties of red fluorescent proteins and provides the basis for rational engineering of this subfamily of GFP homologs.

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Figure 1: Overall structure of DsRed.
Figure 2: The chromophore structure of DsRed.
Figure 3: Stereoview of the DsRed oligomerization interfaces.
Figure 4: Structure-based sequence alignment of fluorescent protein family.

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Acknowledgements

We are grateful to Z. Otwinowski for SeMet data collection and initial processing, H. Lee for help preparing crystallization and heavy-atom soak trials, R. Jain, S. Lockless, and M. Machius for insightful discussions, and J. Remington and R. Tsien for communication of data prior to publication. This work was partially supported by a grant from the Robert A. Welch Foundation to R.R., who is also a recipient of the Burroughs-Wellcome Fund New Investigator Award in the Basic Pharmacological Sciences, and is an Assistant Investigator of the Howard Hughes Medical Institute.

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  1. Howard Hughes Medical Institute and Department of Pharmacology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, 75390-9050, Texas, USA

    Mark A. Wall, Michael Socolich & Rama Ranganathan

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  1. Mark A. Wall
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Correspondence to Rama Ranganathan.

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Wall, M., Socolich, M. & Ranganathan, R. The structural basis for red fluorescence in the tetrameric GFP homolog DsRed. Nat Struct Mol Biol 7, 1133–1138 (2000). https://doi.org/10.1038/81992

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