Structure and mechanism of the photoactivatable green fluorescent protein

J. Nathan Henderson, Rinat Gepshtein, Josef R. Heenan, Karen Kallio, Dan Huppert, S. James Remington

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)4176-4177
Number of pages2
JournalJournal of the American Chemical Society
Volume131
Issue number12
DOIs
StatePublished - 1 Apr 2009

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