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Structure and mechanism of the photoactivatable green fluorescent protein.
J Am Chem Soc. 2009 Apr 01; 131(12):4176-7.JA

Abstract

Crystal structures of the photoactivatable green fluorescent protein T203H variant (PA-GFP) have been solved in the native and photoactivated states, which under 488 nm illumination are dark and brightly fluorescent, respectively. We demonstrate that photoactivation of PA-GFP is the result of a UV-induced decarboxylation of the Glu222 side chain that shifts the chromophore equilibrium to the anionic form. Coupled with the T203H mutation, which stabilizes the native PA-GFP neutral chromophore, Glu222 decarboxylation yields a 100-fold contrast enhancement relative to wild-type GFP (WT). Additionally, the structures provide insights into the spectroscopic differences between WT and PA-GFP steady-state fluorescence maxima and excited-state proton transfer dynamics.

Authors+Show Affiliations

Institute of Molecular Biology and Department of Physics, University of Oregon, Eugene, Oregon 97403-1229, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

19278226

Citation

Henderson, J Nathan, et al. "Structure and Mechanism of the Photoactivatable Green Fluorescent Protein." Journal of the American Chemical Society, vol. 131, no. 12, 2009, pp. 4176-7.
Henderson JN, Gepshtein R, Heenan JR, et al. Structure and mechanism of the photoactivatable green fluorescent protein. J Am Chem Soc. 2009;131(12):4176-7.
Henderson, J. N., Gepshtein, R., Heenan, J. R., Kallio, K., Huppert, D., & Remington, S. J. (2009). Structure and mechanism of the photoactivatable green fluorescent protein. Journal of the American Chemical Society, 131(12), 4176-7. https://doi.org/10.1021/ja808851n
Henderson JN, et al. Structure and Mechanism of the Photoactivatable Green Fluorescent Protein. J Am Chem Soc. 2009 Apr 1;131(12):4176-7. PubMed PMID: 19278226.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structure and mechanism of the photoactivatable green fluorescent protein. AU - Henderson,J Nathan, AU - Gepshtein,Rinat, AU - Heenan,Josef R, AU - Kallio,Karen, AU - Huppert,Dan, AU - Remington,S James, PY - 2009/3/13/entrez PY - 2009/3/13/pubmed PY - 2009/7/29/medline SP - 4176 EP - 7 JF - Journal of the American Chemical Society JO - J Am Chem Soc VL - 131 IS - 12 N2 - Crystal structures of the photoactivatable green fluorescent protein T203H variant (PA-GFP) have been solved in the native and photoactivated states, which under 488 nm illumination are dark and brightly fluorescent, respectively. We demonstrate that photoactivation of PA-GFP is the result of a UV-induced decarboxylation of the Glu222 side chain that shifts the chromophore equilibrium to the anionic form. Coupled with the T203H mutation, which stabilizes the native PA-GFP neutral chromophore, Glu222 decarboxylation yields a 100-fold contrast enhancement relative to wild-type GFP (WT). Additionally, the structures provide insights into the spectroscopic differences between WT and PA-GFP steady-state fluorescence maxima and excited-state proton transfer dynamics. SN - 1520-5126 UR - https://www.unboundmedicine.com/medline/citation/19278226/Structure_and_mechanism_of_the_photoactivatable_green_fluorescent_protein_ L2 - https://doi.org/10.1021/ja808851n DB - PRIME DP - Unbound Medicine ER -