A Tandem Green–Red Heterodimeric Fluorescent Protein with High FRET Efficiency

@article{Wiens2016ATG,
  title={A Tandem Green–Red Heterodimeric Fluorescent Protein with High FRET Efficiency},
  author={Matthew D. Wiens and Yi Shen and Xi Li and M. Alaraby Salem and Nick Smisdom and Wei Zhang and Alex Brown and Robert E. Campbell},
  journal={ChemBioChem},
  year={2016},
  volume={17}
}
The tetrameric red fluorescent protein from Discosoma sp. coral (DsRed) has previously been engineered to produce dimeric and monomeric fluorescent variants with excitation and emission profiles that span the visible spectrum. The brightest of the effectively monomeric DsRed variants is tdTomato—a tandem fusion of a dimeric DsRed variant. Here we describe the engineering of brighter red (RRvT), green (GGvT), and green–red heterodimeric (GRvT) tdTomato variants. GRvT exhibited 99… Expand
Association of Fluorescent Protein Pairs and Its Significant Impact on Fluorescence and Energy Transfer
TLDR
Simple assumptions used when monitoring interactions between proteins via FP FRET may not always hold true, especially under conditions whereby the protein–protein interactions promote FP interaction. Expand
The effect of proximity on the function and energy transfer capability of fluorescent protein pairs
TLDR
It is shown here that on association of FP pairs, the inherent function of the FPs can alter, and the simple assumptions used when monitoring interactions between proteins via FP FRET may not always hold true. Expand
Ratiometric BRET Measurements of ATP with a Genetically-Encoded Luminescent Sensor
TLDR
A new ratiometric BRET sensor of ATP (ARSeNL—ATP detection with a Ratiometric mScarlet-NanoLuc sensor) is generated and demonstrated that it provides a stable and robust readout across protein, cell, and whole animal tissue contexts, and could provide a new tool for in vivo imaging of metabolic status. Expand
Dual-Mode FRET and BRET Sensors for Detecting cAMP Dynamics
TLDR
This work engineered an Epac1-based sensor that responds to cyclic adenosine monophosphate binding with a greater than 80% change in both Förster Resonance Energy Transfer and bioluminescent resonance energy transfer modes. Expand
DNA binding fluorescent proteins as single-molecule probes.
TLDR
The design of FP-DBPs is summarized and their brightness, photostability, pKa, maturation rate, and binding affinity (Kd) characteristics are reviewed, and single DNA molecule visualisation using FP- DBPs is focused on. Expand
Quantitative single-molecule imaging of protein assembly in membranes
Abstract Accurate knowledge of the mechanisms of assembly of protein complexes is essential to understand their structure and control their function. Deep insight into these mechanisms requiresExpand
Anterograde Neuronal Circuit Tracers Derived from Herpes Simplex Virus 1: Development, Application, and Perspectives
TLDR
This review focuses on the current H129-derived viral tracers and highlights strategies in which future technological development can advance HSV-1's use as a tool. Expand
HIV-1 uncoats in the nucleus near sites of integration
TLDR
The results show that intact (or nearly intact) viral cores enter the nucleus through a mechanism involving interactions with host protein cleavage and polyadenylation specificity factor 6 (CPSF6), complete reverse transcription in the nucleus before uncoating, and uncoat near their integration sites just before integration. Expand

References

SHOWING 1-10 OF 43 REFERENCES
An engineered monomeric Zoanthus sp. yellow fluorescent protein.
TLDR
It is demonstrated that mPapaya1 can serve as a good Förster resonance energy transfer (FRET) acceptor when paired with an mTFP1 donor and is a valuable addition to the palette of FP variants that are useful for multicolor imaging and FRET-based biosensing. Expand
Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein
TLDR
The latest red version matures more completely, is more tolerant of N-terminal fusions and is over tenfold more photostable than mRFP1, and three monomers with distinguishable hues from yellow-orange to red-orange have higher quantum efficiencies. Expand
Dimerization-dependent green and yellow fluorescent proteins.
TLDR
Using the ddGFP tool, this work generated a highly effective indicator of endomembrane proximity that can be used to image the mitochondria-associated membrane (MAM) interface of endoplasmic reticulum (ER) and mitochondria. Expand
A monomeric red fluorescent protein
  • R. Campbell, Oded Tour, +4 authors R. Tsien
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 2002
TLDR
This work presents the stepwise evolution of DsRed to a dimer and then either to a genetic fusion of two copies of the protein, i.e., a tandem dimer, or to a true monomer designated mRFP1 (monomeric red fluorescent protein). Expand
Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed)
TLDR
To overcome the slow maturation, random and directed mutagenesis is used to create DsRed variants that mature 10–15 times faster than the wild-type protein and yield bright fluorescence even in rapidly growing organisms such as yeast. Expand
Improving FRET dynamic range with bright green and red fluorescent proteins
TLDR
Replacement of CFP and YFP with these two proteins in reporters of kinase activity, small GTPase activity and transmembrane voltage significantly improves photostability, FRET dynamic range and emission ratio changes and enhances detection of transient biochemical events. Expand
GFP‐like chromoproteins as a source of far‐red fluorescent proteins
TLDR
A dimeric mutant with similar maturation and spectral properties to tetrameric HcRed, which is excitable by 600 nm dye laser, thus promoting new detection channels for multi‐color flow cytometry applications. Expand
Robust red FRET sensors using self-associating fluorescent domains.
TLDR
A generic strategy to enhance the traditionally poor sensitivity of red FRET sensors by developing self-associating variants of mOrange and mCherry that allow sensors to switch between well-defined on- and off states is presented. Expand
Evolutionary optimization of fluorescent proteins for intracellular FRET
TLDR
A quantitative evolutionary strategy using fluorescence-activated cell sorting to optimize a cyan-yellow fluorescent protein pair for FRET provides substantially improved sensitivity and dynamic range for a broad range of molecular imaging and screening applications. Expand
The nature of fluorescence emission in the red fluorescent protein DsRed, revealed by single-molecule detection
TLDR
The results indicate that energy transfer between identical monomers occurs efficiently with red emission arising equally likely from any of the chromophoric units, and oligomer suppression would not only be advantageous for protein fusion but will also increase the fluorescence emission of individual monomers. Expand
...
1
2
3
4
5
...