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Contents list

Analysis of omics data with genome-scale models of metabolism

Protein co-evolution: how do we combine bioinformatics and experimental approaches?

Systematic analysis of histone modification readout

Early self-reproduction, the emergence of division mechanisms in protocells

Reconstruction and analysis of the genome-scale metabolic network of Candida glabrata

Dissection of the potential characteristic of miRNA–miRNA functional synergistic regulations

From time series to biological network regulations: an evolutionary approach

Biochemical measurements on single erythroid progenitor cells shed light on the combinatorial regulation of red blood cell production

Role of block copolymer-micelle nanocomposites in dye–bovine serum albumin binding: a combined experimental and molecular docking study

Minimum set of mutations needed to optimize cyan fluorescent proteins for live cell imaging

Dynamic and structural constraints in signal propagation by regulatory networks

Systems-level characterization and engineering of oxidative stress tolerance in Escherichia coli under anaerobic conditions

A cellular stress-directed bistable switch controls the crosstalk between autophagy and apoptosis

Quantitative metabolomic and lipidomic profiling reveals aberrant amino acid metabolism in type 2 diabetes

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Minimum set of mutations needed to optimize cyan fluorescent proteins for live cell imaging

Content Provider	Royal Society of Chemistry (RSC)
Author	Erard, Marie Bousmah, Yasmina Fredj, Asma Derrien, Valérie Pasquier, Hélène Beltolngar, Dahdjim-Benoît Merola, Fabienne Vincent, Pierre
Copyright Year	2013
Abstract	Cyan fluorescent proteins (CFPs) are widely used as FRET donors in genetically encoded biosensors for live cell imaging. Recently, cyan variants with greatly improved fluorescence quantum yields have been developed by large scale random mutagenesis. We show that the introduction of only two mutations, T65S and H148G, is able to confer equivalent performances on the popular form ECFP, leading to Aquamarine (QY = 0.89, τf = 4.12 ns). Besides an impressive pH stability (pK1/2 = 3.3), Aquamarine shows a very low general sensitivity to its environment, and undetectable photoswitching reactions. Aquamarine gives efficient and bright expression in different mammalian cell systems, with a long and single exponential intracellular fluorescence lifetime mostly insensitive to the fusion or the subcellular location of the protein. Aquamarine was also able to advantageously replace the CFP donor in the FRET biosensor AKAR for ratiometric measurements of protein kinase A activity. The performances of Aquamarine show that only two rounds of straightforward single point mutagenesis can be a quick and efficient way to optimize the donor properties in FRET-based biosensors.
Starting Page	258
Ending Page	267
Page Count	10
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	9
Issue Number	2
Journal	Molecular BioSystems
DOI	10.1039/c2mb25303h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Protein kinase A Förster resonance energy transfer Cyan Beryl AKAR Biosensor Fluorescence PH Quantum information science Treatment as prevention CFP Protein
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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Monitoring Agonist-induced Phospholipase C Activation in Live Cells by Fluorescence Resonance Energy Transfer

Simultaneous real-time detection of initiator- and effector-caspase activation by double fluorescence resonance energy transfer analysis.

Optimization of Cyan Fluorescent Protein Fluorescence for Förster Resonance Energy Transfer

Molecular Modeling of Fluorescent SERCA Biosensors.

Visualization of phosphatase activity in living cells with a FRET-based calcineurin activity sensor

tdLanYFP, a Yellow, Bright, Photostable, and pH-Insensitive Fluorescent Protein for Live-Cell Imaging and Förster Resonance Energy Transfer-Based Sensing Strategies

Genetically encoded fluorescent reporters of histone methylation in living cells.

Some Secrets of Fluorescent Proteins: Distinct Bleaching in Various Mounting Fluids and Photoactivation of Cyan Fluorescent Proteins at YFP-Excitation

Atp changes the fluorescence lifetime of cyan fluorescent protein via an interaction with his148 (2010).

Minimum set of mutations needed to optimize cyan fluorescent proteins for live cell imaging

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Monitoring Agonist-induced Phospholipase C Activation in Live Cells by Fluorescence Resonance Energy Transfer

Simultaneous real-time detection of initiator- and effector-caspase activation by double fluorescence resonance energy transfer analysis.

Optimization of Cyan Fluorescent Protein Fluorescence for Förster Resonance Energy Transfer

Molecular Modeling of Fluorescent SERCA Biosensors.

Visualization of phosphatase activity in living cells with a FRET-based calcineurin activity sensor

tdLanYFP, a Yellow, Bright, Photostable, and pH-Insensitive Fluorescent Protein for Live-Cell Imaging and Förster Resonance Energy Transfer-Based Sensing Strategies

Genetically encoded fluorescent reporters of histone methylation in living cells.

Some Secrets of Fluorescent Proteins: Distinct Bleaching in Various Mounting Fluids and Photoactivation of Cyan Fluorescent Proteins at YFP-Excitation

Atp changes the fluorescence lifetime of cyan fluorescent protein via an interaction with his148 (2010).

Minimum set of mutations needed to optimize cyan fluorescent proteins for live cell imaging