scholarly journals A Green Fluorescent Protein Containing a QFG Tri-Peptide Chromophore: Optical Properties and X-Ray Crystal Structure

PLoS ONE ◽  
2012 ◽  
Vol 7 (10) ◽  
pp. e47331 ◽  
Author(s):  
Jion M. Battad ◽  
Daouda A. K. Traore ◽  
Emma Byres ◽  
Jamie Rossjohn ◽  
Rodney J. Devenish ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Optical Properties ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
X Ray ◽  
Green Fluorescent

scholarly journals The structure of a GFP-based antibody (fluorobody) to TLH, a toxin fromVibrio parahaemolyticus

2015 ◽  
Vol 71 (7) ◽  
pp. 913-918 ◽  
Author(s):  
Yaoguang Chen ◽  
Xiaocheng Huang ◽  
Rongzhi Wang ◽  
Shihua Wang ◽  
Ning Shi
Keyword(s):  
Crystal Structure ◽  
Green Fluorescent Protein ◽  
Vibrio Parahaemolyticus ◽  
Fluorescent Protein ◽  
Hybrid Molecule ◽  
Unit Cell Parameters ◽  
Cell Parameters ◽  
Food Borne ◽  
Vapour Diffusion ◽  
Green Fluorescent

A fluorobody is a manmade hybrid molecule that is composed of green fluorescent protein (GFP) and a fragment of antibody, which combines the affinity and specificity of an antibody with the visibility of a GFP. It is able to provide a real-time indication of binding while avoiding the use of tags and secondary binding reagents. Here, the expression, purification and crystal structure of a recombinant fluorobody for TLH (thermolabile haemolysin), a toxin from the lethal food-borne disease bacteriumVibrio parahaemolyticus, are presented. This is the first structure of a fluorobody to be reported. Crystals belonging to space groupP43212, with unit-cell parametersa=b= 63.35,c = 125.90 Å, were obtained by vapour diffusion in hanging drops and the structure was refined to anRfreeof 16.7% at 1.5 Å resolution. The structure shows a CDR loop of the antibody on the GFP scaffold.

scholarly journals Nonlinear Optical Properties of Green Fluorescent Protein Chromophore Coupled Diradicals

2013 ◽  
Vol 29 (12) ◽  
pp. 2543-2550
Author(s):  
YU Hai-Ling ◽  
◽  
ZHANG Meng-Ying ◽  
HONG Bo ◽  
CHENG Zhi Qiang ◽  
...  
Keyword(s):  
Optical Properties ◽  
Green Fluorescent Protein ◽  
Nonlinear Optical ◽  
Fluorescent Protein ◽  
Nonlinear Optical Properties ◽  
Green Fluorescent

scholarly journals X-ray crystallographic studies on the hydrogen isotope effects of green fluorescent protein at sub-ångström resolutions

2019 ◽  
Vol 75 (12) ◽  
pp. 1096-1106 ◽  
Author(s):  
Yang Tai ◽  
Kiyofumi Takaba ◽  
Yuya Hanazono ◽  
Hoang-Anh Dao ◽  
Kunio Miki ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Active Site ◽  
Hydrogen Isotope ◽  
Fluorescent Protein ◽  
Isotope Effects ◽  
Protonation State ◽  
X Ray ◽  
Two Samples ◽  
Deuterated Proteins ◽  
Green Fluorescent

Hydrogen atoms are critical to the nature and properties of proteins, and thus deuteration has the potential to influence protein function. In fact, it has been reported that some deuterated proteins show different physical and chemical properties to their protiated counterparts. Consequently, it is important to investigate protonation states around the active site when using deuterated proteins. Here, hydrogen isotope effects on the S65T/F99S/M153T/V163A variant of green fluorescent protein (GFP), in which the deprotonated B form is dominant at pH 8.5, were investigated. The pH/pD dependence of the absorption and fluorescence spectra indicates that the protonation state of the chromophore is the same in protiated GFP in H2O and protiated GFP in D2O at pH/pD 8.5, while the pK a of the chromophore became higher in D2O. Indeed, X-ray crystallographic analyses at sub-ångström resolution revealed no apparent changes in the protonation state of the chromophore between the two samples. However, detailed comparisons of the hydrogen OMIT maps revealed that the protonation state of His148 in the vicinity of the chromophore differed between the two samples. This indicates that protonation states around the active site should be carefully adjusted to be the same as those of the protiated protein when neutron crystallographic analyses of proteins are performed.

Expression and optical properties of green fluorescent protein expressed in different cellular environments

2005 ◽  
Vol 119 (4) ◽  
pp. 368-378 ◽  
Author(s):  
Jin Zou ◽  
Yiming Ye ◽  
Kristy Welshhans ◽  
Monica Lurtz ◽  
April L. Ellis ◽  
...  
Keyword(s):  
Optical Properties ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Green Fluorescent

scholarly journals Structural analysis of the bright monomeric yellow-green fluorescent protein mNeonGreen obtained by directed evolution

2016 ◽  
Vol 72 (12) ◽  
pp. 1298-1307 ◽  
Author(s):  
Damien Clavel ◽  
Guillaume Gotthard ◽  
David von Stetten ◽  
Daniele De Sanctis ◽  
Hélène Pasquier ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Directed Evolution ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Yellow Fluorescent Protein ◽  
X Rays ◽  
Visible Absorption ◽  
X Ray ◽  
X Ray Crystallography ◽  
Green Fluorescent

Until recently, genes coding for homologues of the autofluorescent protein GFP had only been identified in marine organisms from the phyla Cnidaria and Arthropoda. New fluorescent-protein genes have now been found in the phylum Chordata, coding for particularly bright oligomeric fluorescent proteins such as the tetrameric yellow fluorescent proteinlanYFP fromBranchiostoma lanceolatum. A successful monomerization attempt led to the development of the bright yellow-green fluorescent protein mNeonGreen. The structures oflanYFP and mNeonGreen have been determined and compared in order to rationalize the directed evolution process leading from a bright, tetrameric to a still bright, monomeric fluorescent protein. An unusual discolouration of crystals of mNeonGreen was observed after X-ray data collection, which was investigated using a combination of X-ray crystallography and UV–visible absorption and Raman spectroscopies, revealing the effects of specific radiation damage in the chromophore cavity. It is shown that X-rays rapidly lead to the protonation of the phenolate O atom of the chromophore and to the loss of its planarity at the methylene bridge.

Crystal Structure and Photodynamic Behavior of the Blue Emission Variant Y66H/Y145F of Green Fluorescent Protein†

Biochemistry ◽  
1997 ◽  
Vol 36 (32) ◽  
pp. 9759-9765 ◽  
Author(s):  
Rebekka M. Wachter ◽  
Brett A. King ◽  
Roger Heim ◽  
Karen Kallio ◽  
Roger Y. Tsien ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Blue Emission ◽  
Green Fluorescent

scholarly journals Dynamic tuning of FRET in a green fluorescent protein biosensor

2019 ◽  
Vol 5 (8) ◽  
pp. eaaw4988 ◽  
Author(s):  
Pablo Trigo-Mourino ◽  
Thomas Thestrup ◽  
Oliver Griesbeck ◽  
Christian Griesinger ◽  
Stefan Becker
Keyword(s):  
Green Fluorescent Protein ◽  
Protein Interactions ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Structural Data ◽  
X Ray ◽  
Dynamic Tuning ◽  
Green Fluorescent

Förster resonance energy transfer (FRET) between mutants of green fluorescent protein is widely used to monitor protein-protein interactions and as a readout mode in fluorescent biosensors. Despite the fundamental importance of distance and molecular angles of fluorophores to each other, structural details on fluorescent protein FRET have been missing. Here, we report the high-resolution x-ray structure of the fluorescent proteins mCerulean3 and cpVenus within the biosensor Twitch-2B, as they undergo FRET and characterize the dynamics of this biosensor with B02-dependent paramagnetic nuclear magnetic resonance at 900 MHz and 1.1 GHz. These structural data provide the unprecedented opportunity to calculate FRET from the x-ray structure and to compare it to experimental data in solution. We find that interdomain dynamics limits the FRET effect and show that a rigidification of the sensor further enhances FRET.

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