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The 1.7 A crystal structure of Dronpa: a photoswitchable green fluorescent protein.
J Mol Biol. 2006 Nov 24; 364(2):213-24.JM

Abstract

The green fluorescent protein (GFP), its variants, and the closely related GFP-like proteins possess a wide variety of spectral properties that are of widespread interest as biological tools. One desirable spectral property, termed photoswitching, involves the light-induced alteration of the optical properties of certain GFP members. Although the structural basis of both reversible and irreversible photoswitching events have begun to be unraveled, the mechanisms resulting in reversible photoswitching are less clear. A novel GFP-like protein, Dronpa, was identified to have remarkable light-induced photoswitching properties, maintaining an almost perfect reversible photochromic behavior with a high fluorescence to dark state ratio. We have crystallized and subsequently determined to 1.7 A resolution the crystal structure of the fluorescent state of Dronpa. The chromophore was observed to be in its anionic form, adopting a cis co-planar conformation. Comparative structural analysis of non-photoactivatable and photoactivatable GFPs, together with site-directed mutagenesis of a position (Cys62) within the Dronpa chromophore, has provided a basis for understanding Dronpa photoactivation. Specifically, we propose a model of reversible photoactivation whereby irradiation with light leads to subtle conformational changes within and around the environment of the chromophore that promotes proton transfer along an intricate polar network.

Authors+Show Affiliations

The Protein Crystallography Unit, Monash University, Clayton, Victoria 3800, Australia.No affiliation info availableNo 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

Language

eng

PubMed ID

17010376

Citation

Wilmann, Pascal G., et al. "The 1.7 a Crystal Structure of Dronpa: a Photoswitchable Green Fluorescent Protein." Journal of Molecular Biology, vol. 364, no. 2, 2006, pp. 213-24.
Wilmann PG, Turcic K, Battad JM, et al. The 1.7 A crystal structure of Dronpa: a photoswitchable green fluorescent protein. J Mol Biol. 2006;364(2):213-24.
Wilmann, P. G., Turcic, K., Battad, J. M., Wilce, M. C., Devenish, R. J., Prescott, M., & Rossjohn, J. (2006). The 1.7 A crystal structure of Dronpa: a photoswitchable green fluorescent protein. Journal of Molecular Biology, 364(2), 213-24.
Wilmann PG, et al. The 1.7 a Crystal Structure of Dronpa: a Photoswitchable Green Fluorescent Protein. J Mol Biol. 2006 Nov 24;364(2):213-24. PubMed PMID: 17010376.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The 1.7 A crystal structure of Dronpa: a photoswitchable green fluorescent protein. AU - Wilmann,Pascal G, AU - Turcic,Kristina, AU - Battad,Jion M, AU - Wilce,Matthew C J, AU - Devenish,Rodney J, AU - Prescott,Mark, AU - Rossjohn,Jamie, Y1 - 2006/09/03/ PY - 2006/06/07/received PY - 2006/08/24/revised PY - 2006/08/31/accepted PY - 2006/10/3/pubmed PY - 2007/1/11/medline PY - 2006/10/3/entrez SP - 213 EP - 24 JF - Journal of molecular biology JO - J Mol Biol VL - 364 IS - 2 N2 - The green fluorescent protein (GFP), its variants, and the closely related GFP-like proteins possess a wide variety of spectral properties that are of widespread interest as biological tools. One desirable spectral property, termed photoswitching, involves the light-induced alteration of the optical properties of certain GFP members. Although the structural basis of both reversible and irreversible photoswitching events have begun to be unraveled, the mechanisms resulting in reversible photoswitching are less clear. A novel GFP-like protein, Dronpa, was identified to have remarkable light-induced photoswitching properties, maintaining an almost perfect reversible photochromic behavior with a high fluorescence to dark state ratio. We have crystallized and subsequently determined to 1.7 A resolution the crystal structure of the fluorescent state of Dronpa. The chromophore was observed to be in its anionic form, adopting a cis co-planar conformation. Comparative structural analysis of non-photoactivatable and photoactivatable GFPs, together with site-directed mutagenesis of a position (Cys62) within the Dronpa chromophore, has provided a basis for understanding Dronpa photoactivation. Specifically, we propose a model of reversible photoactivation whereby irradiation with light leads to subtle conformational changes within and around the environment of the chromophore that promotes proton transfer along an intricate polar network. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/17010376/The_1_7_A_crystal_structure_of_Dronpa:_a_photoswitchable_green_fluorescent_protein_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(06)01166-1 DB - PRIME DP - Unbound Medicine ER -