scholarly journals Identification of mNeonGreen as a pH‐Dependent, Turn‐On Fluorescent Protein Sensor for Chloride

ChemBioChem ◽  
2019 ◽  
Author(s):  
Jasmine N. Tutol ◽  
Hiu C. Kam ◽  
Sheel C. Dodani
Keyword(s):  
Fluorescent Protein ◽  
Turn On ◽  
Ph Dependent ◽  
Protein Sensor

scholarly journals The fluorescent protein sensor roGFP2‐Orp1 monitorsin vivoH2O2and thiol redox integration and elucidates intracellular H2O2dynamics during elicitor‐induced oxidative burst in Arabidopsis

2018 ◽  
Vol 221 (3) ◽  
pp. 1649-1664 ◽  
Author(s):  
Thomas Nietzel ◽  
Marlene Elsässer ◽  
Cristina Ruberti ◽  
Janina Steinbeck ◽  
José Manuel Ugalde ◽  
...  
Keyword(s):  
Oxidative Burst ◽  
Fluorescent Protein ◽  
Thiol Redox ◽  
Protein Sensor

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.

pH-Dependent Fluorescence of a Heterologously ExpressedAequoreaGreen Fluorescent Protein Mutant: In Situ Spectral Characteristics and Applicability to Intracellular pH Estimation†,‡

Biochemistry ◽  
1998 ◽  
Vol 37 (28) ◽  
pp. 9894-9901 ◽  
Author(s):  
R. Brooks Robey ◽  
Ofelia Ruiz ◽  
Anna V. P. Santos ◽  
Jianfei Ma ◽  
Fely Kear ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Fluorescent Protein ◽  
Spectral Characteristics ◽  
Ph Dependent

scholarly journals Lysozyme coated copper nanoclusters for green fluorescence and their utility in cell imaging

2020 ◽  
Vol 1 (5) ◽  
pp. 1439-1447
Author(s):  
Atul Gajanan Thawari ◽  
Piyush Kumar ◽  
Rohit Srivastava ◽  
Chebrolu Pulla Rao
Keyword(s):  
Green Fluorescent Protein ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cell Imaging ◽  
Fluorescent Protein ◽  
Water Soluble ◽  
Green Fluorescence ◽  
Copper Nanoclusters ◽  
Ph Dependent ◽  
Green Fluorescent

Green fluorescent, pH dependent and water soluble copper nanoclusters (3–5 nm) were synthesized by stabilizing with lysozyme and these were demonstrated for imaging in both healthy and cancer cell lines as an alternate to green fluorescent protein.

Live pancreatic acinar imaging of exocytosis using syncollin-pHluorin

2011 ◽  
Vol 300 (6) ◽  
pp. C1513-C1523 ◽  
Author(s):  
Nestor A. Fernandez ◽  
Tao Liang ◽  
Herbert Y. Gaisano
Keyword(s):  
Fusion Protein ◽  
Protein Expression ◽  
Fluorescent Protein ◽  
Acinar Cells ◽  
Three Dimensions ◽  
Imaging Method ◽  
Zymogen Granule ◽  
Ph Dependent ◽  
Green Fluorescent ◽  
Superior Approach

In this report, a novel live acinar exocytosis imaging technique is described. An adenovirus was engineered, encoding for an endogenous zymogen granule (ZG) protein (syncollin) fused to pHluorin, a pH-dependent green fluorescent protein (GFP). Short-term culture of mouse acini infected with this virus permits exogenous adenoviral protein expression while retaining acinar secretory competence and cell polarity. The syncollin-pHluorin fusion protein was shown to be correctly localized to ZGs, and the pH-dependent fluorescence of pHluorin was retained. Coupled with the use of a spinning disk confocal microscope, the syncollin-pHluorin fusion protein exploits the ZG luminal pH changes that occur during exocytosis to visualize exocytic events of live acinar cells in real-time with high spatial resolution in three dimensions. Apical and basolateral exocytic events were observed on stimulation of acinar cells with maximal and supramaximal cholecystokinin concentrations, respectively. Sequential exocytic events were also observed. Coupled with the use of transgenic mice and/or adenovirus-mediated protein expression, this syncollin-pHluorin imaging method offers a superior approach to studying pancreatic acinar exocytosis. This assay can also be applied to acinar disease models to elucidate the mechanisms implicated in pancreatitis.

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