Tags

Type your tag names separated by a space and hit enter

Structural basis of enhanced photoconversion yield in green fluorescent protein-like protein Dendra2.
Biochemistry. 2009 Jun 09; 48(22):4905-15.B

Abstract

Dendra2 is an engineered, monomeric GFP-like protein that belongs to a subclass of fluorescent proteins undergoing irreversible photoconversion from a green- to a red-emitting state upon exposure to purple-blue light. This photoinduced process occurs only in the neutral state of the chromophore and is known to result from backbone cleavage accompanied by an extension of the delocalized pi-electron system. We have measured the X-ray structure of the green species of Dendra2 and performed a comprehensive characterization of the optical absorption and fluorescence properties of the protein in both its green and red forms. The structure, which is very similar to those reported for the closely related proteins EosFP and Kaede, revealed a local structural change involving mainly Arg66 and a water molecule W4, which are part of a charged and hydrogen-bonded cluster of amino acids and water molecules next to the chromophore. Unlike in EosFP and Kaede, Arg66 of Dendra2 does not contribute to negative charge stabilization on the imidazolinone ring by hydrogen bonding to the imidazolinone carbonyl. This structural change may explain the blue shift of the absorption and emission bands, as well as the markedly higher pKs of the hydroxyphenyl moiety of the chromophore, which were determined as 7.1 and 7.5 for the green and red species, respectively. The action spectrum of photoconversion coincides with the absorption band of the neutral species. Consequently, its 20-fold enhancement in Dendra2 at physiological pH accounts for the higher photoconversion yield of this protein as compared to EosFP.

Authors+Show Affiliations

European Synchrotron Radiation Facility, 6 Rue Jules Horowitz, BP 220, 8043 Grenoble Cedex, France.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19371086

Citation

Adam, Virgile, et al. "Structural Basis of Enhanced Photoconversion Yield in Green Fluorescent Protein-like Protein Dendra2." Biochemistry, vol. 48, no. 22, 2009, pp. 4905-15.
Adam V, Nienhaus K, Bourgeois D, et al. Structural basis of enhanced photoconversion yield in green fluorescent protein-like protein Dendra2. Biochemistry. 2009;48(22):4905-15.
Adam, V., Nienhaus, K., Bourgeois, D., & Nienhaus, G. U. (2009). Structural basis of enhanced photoconversion yield in green fluorescent protein-like protein Dendra2. Biochemistry, 48(22), 4905-15. https://doi.org/10.1021/bi900383a
Adam V, et al. Structural Basis of Enhanced Photoconversion Yield in Green Fluorescent Protein-like Protein Dendra2. Biochemistry. 2009 Jun 9;48(22):4905-15. PubMed PMID: 19371086.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structural basis of enhanced photoconversion yield in green fluorescent protein-like protein Dendra2. AU - Adam,Virgile, AU - Nienhaus,Karin, AU - Bourgeois,Dominique, AU - Nienhaus,G Ulrich, PY - 2009/4/18/entrez PY - 2009/4/18/pubmed PY - 2009/7/23/medline SP - 4905 EP - 15 JF - Biochemistry JO - Biochemistry VL - 48 IS - 22 N2 - Dendra2 is an engineered, monomeric GFP-like protein that belongs to a subclass of fluorescent proteins undergoing irreversible photoconversion from a green- to a red-emitting state upon exposure to purple-blue light. This photoinduced process occurs only in the neutral state of the chromophore and is known to result from backbone cleavage accompanied by an extension of the delocalized pi-electron system. We have measured the X-ray structure of the green species of Dendra2 and performed a comprehensive characterization of the optical absorption and fluorescence properties of the protein in both its green and red forms. The structure, which is very similar to those reported for the closely related proteins EosFP and Kaede, revealed a local structural change involving mainly Arg66 and a water molecule W4, which are part of a charged and hydrogen-bonded cluster of amino acids and water molecules next to the chromophore. Unlike in EosFP and Kaede, Arg66 of Dendra2 does not contribute to negative charge stabilization on the imidazolinone ring by hydrogen bonding to the imidazolinone carbonyl. This structural change may explain the blue shift of the absorption and emission bands, as well as the markedly higher pKs of the hydroxyphenyl moiety of the chromophore, which were determined as 7.1 and 7.5 for the green and red species, respectively. The action spectrum of photoconversion coincides with the absorption band of the neutral species. Consequently, its 20-fold enhancement in Dendra2 at physiological pH accounts for the higher photoconversion yield of this protein as compared to EosFP. SN - 1520-4995 UR - https://www.unboundmedicine.com/medline/citation/19371086/Structural_basis_of_enhanced_photoconversion_yield_in_green_fluorescent_protein_like_protein_Dendra2_ L2 - https://dx.doi.org/10.1021/bi900383a DB - PRIME DP - Unbound Medicine ER -