scholarly journals A general approach to engineer positive-going eFRET voltage indicators

2019 ◽  
Author(s):  
Ahmed S. Abdelfattah ◽  
Rosario Valenti ◽  
Allan Wong ◽  
Minoru Koyama ◽  
Douglas S. Kim ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Proton Transport ◽  
Fluorescent Protein ◽  
State Of The Art ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Fluorescence Signal ◽  
Transport Pathway ◽  
Signal Polarity

We engineered electrochromic fluorescence resonance energy transfer (eFRET) genetically encoded voltage indicators (GEVIs) with “positive-going” fluorescence response to membrane depolarization through rational manipulation of the native proton transport pathway in microbial rhodopsins. We transformed the state-of-the-art eFRET GEVI Voltron into Positron, with kinetics and sensitivity equivalent to Voltron but flipped fluorescence signal polarity. We further applied this general approach to GEVIs containing different voltage sensitive rhodopsin domains and various fluorescent dye and fluorescent protein reporters.

scholarly journals Physical Association between the Adipocyte Fatty Acid-binding Protein and Hormone-sensitive Lipase

2004 ◽  
Vol 279 (50) ◽  
pp. 52399-52405 ◽  
Author(s):  
Anne J. Smith ◽  
Mark A. Sanders ◽  
Brian R. Thompson ◽  
Constantine Londos ◽  
Fredric B. Kraemer ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Fatty Acid ◽  
Fluorescence Resonance Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Hormone Sensitive Lipase ◽  
Fatty Acid Binding ◽  
Hormone Sensitive ◽  
Fluorescence Resonance

Previousin vitrostudies have established that hormone sensitive lipase (HSL) and adipocyte fatty acid-binding protein (AFABP) form a physical complex that presumably positions the FABP to accept a product fatty acid generated during catalysis. To assess AFABP-HSL interaction within a cellular context, we have used lipocytes derived from 293 cells (C8PA cells) and examined physical association using fluorescence resonance energy transfer. Transfection of C8PA cells with cyan fluorescent protein (CFP)-HSL, yellow fluorescent protein (YFP)-adipocyte FABP, or YFP-liver FABP revealed that under basal conditions each protein was cytoplasmic. In the presence of 20 μmforskolin, CFP-HSL translocated to the triacylglycerol droplet, coincident with BODIPY-FA labeled depots. Fluorescence resonance energy transfer analysis demonstrated that CFP-HSL associated with YFP-adipocyte FABP in both basal and forskolin-treated cells. In contrast, little if any fluorescence resonance energy transfer could be detected between CFP-HSL and YFP-liver FABP. These results suggest that a pre-lipolysis complex containing at least AFABP and HSL exists and that the complex translocates to the surface of the lipid droplet.

scholarly journals Ultra-Sensitive Hydrogen Peroxide Sensor Based on Peroxiredoxin and Fluorescence Resonance Energy Transfer

2020 ◽  
Vol 10 (10) ◽  
pp. 3508
Author(s):  
Haijun Yu ◽  
Haoxiang Li ◽  
Yao Zhou ◽  
Shengmin Zhou ◽  
Ping Wang
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Disulfide Bonds ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Mixed Disulfide ◽  
Fluorescence Resonance ◽  
Protein Sensor

In this paper, a fluorescence resonance energy transfer (FRET)-based sensor for ultra-sensitive detection of H2O2 was developed by utilizing the unique enzymatic properties of peroxiredoxin (Prx) to H2O2. Cyan and yellow fluorescent protein (CFP and YFP) were fused to Prx and mutant thioredoxin (mTrx), respectively. In the presence of H2O2, Prx was oxidized into covalent homodimer through disulfide bonds, which were further reduced by mTrx to form a stable mixed disulfide bond intermediate between CFP-Prx and mTrx-YFP, inducing FRET. A linear quantification range of 10–320 nM was obtained according to the applied protein concentrations and the detection limit (LOD) was determined to be as low as 4 nM. By the assistance of glucose oxidase to transform glucose into H2O2, the CFP-Prx/mTrx-YFP system (CPmTY) was further exploited for the detection of glucose in real sample with good performance, suggesting this CPmTY protein sensor is highly practical.

scholarly journals Interdomain Fluorescence Resonance Energy Transfer in SERCA Probed by Cyan-Fluorescent Protein Fused to the Actuator Domain†

Biochemistry ◽  
2008 ◽  
Vol 47 (14) ◽  
pp. 4246-4256 ◽  
Author(s):  
Deborah L. Winters ◽  
Joseph M. Autry ◽  
Bengt Svensson ◽  
David D. Thomas
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Cyan Fluorescent Protein ◽  
Fluorescence Resonance

Bidirectional Fluorescence Resonance Energy Transfer (FRET) in Mutated and Chemically Modified Yellow Fluorescent Protein (YFP)

2011 ◽  
Vol 22 (2) ◽  
pp. 227-234 ◽  
Author(s):  
Bobin George Abraham ◽  
Nikolai V. Tkachenko ◽  
Ville Santala ◽  
Helge Lemmetyinen ◽  
Matti Karp
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Chemically Modified ◽  
Fluorescence Resonance

Red Fluorescent Protein fromDiscosomaas a Fusion Tag and a Partner for Fluorescence Resonance Energy Transfer†

Biochemistry ◽  
2001 ◽  
Vol 40 (8) ◽  
pp. 2502-2510 ◽  
Author(s):  
Hideaki Mizuno ◽  
Asako Sawano ◽  
Pharhad Eli ◽  
Hiroshi Hama ◽  
Atsushi Miyawaki
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Red Fluorescent Protein ◽  
Fusion Tag ◽  
Fluorescence Resonance

Fluorescence Resonance Energy Transfer-Based Assay for DNA-Binding Protein Tagged by Green Fluorescent Protein

2006 ◽  
Vol 70 (8) ◽  
pp. 1921-1927 ◽  
Author(s):  
Takashi AOKI ◽  
Tomoko IMAMURA ◽  
Hiroyuki OZAKI ◽  
Hideki IDEUCHI ◽  
Shirou TSUCHIDA ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Green Fluorescent Protein ◽  
Dna Binding ◽  
Fluorescence Resonance Energy Transfer ◽  
Fluorescent Protein ◽  
Binding Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Dna Binding Protein ◽  
Green Fluorescent
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