A fluorescence-resonance-energy-transfer-based protease activity assay and its use to monitor paralog-specific small ubiquitin-like modifier processing

2007 ◽  
Vol 363 (1) ◽  
pp. 83-90 ◽  
Author(s):  
Sarah F. Martin ◽  
Neil Hattersley ◽  
Ifor D.W. Samuel ◽  
Ronald T. Hay ◽  
Michael H. Tatham
Keyword(s):  
Energy Transfer ◽  
Fluorescence Resonance Energy Transfer ◽  
Protease Activity ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Activity Assay ◽  
Fluorescence Resonance

scholarly journals Fluorescence Resonance Energy Transfer-Based Assay for Characterization of Hepatitis C Virus NS3-4A Protease Activity in Live Cells

2008 ◽  
Vol 53 (2) ◽  
pp. 728-734 ◽  
Author(s):  
Rosario Sabariegos ◽  
Fernando Picazo ◽  
Beatriz Domingo ◽  
Sandra Franco ◽  
Miguel-Angel Martinez ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Fluorescence Resonance Energy Transfer ◽  
Protease Activity ◽  
Cleavage Site ◽  
Mammalian Cells ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Fluorescence Resonance

ABSTRACT The NS3/4A protease from hepatitis C virus (HCV) plays a key role in viral replication. We report a system for monitoring the activity of this enzyme in single living mammalian cells. We constructed a fluorescence resonance energy transfer (FRET) probe that consists of an enhanced cyan fluorescent protein-citrine fusion, with a cleavage site for HCV NS3/4A protease embedded within the linker between them. Expression of the biosensor in mammalian cells resulted in a FRET signal, and cotransfection with the NS3/4A expression vector produced a significant reduction in FRET, indicating that the cleavage site was processed. Western blot and spectrofluorimetry analysis confirmed the physical cleavage of the fusion probe by the NS3/4A protease. As the level of FRET decay was a function of the protease activity, the system allowed testing of NS3/4A protease variants with different catalytic efficiencies. This FRET probe could be adapted for high-throughput screening of new HCV NS3/4 protease inhibitors.

A multifunctional DNA nanostructure based on multicolor FRET for nuclease activity assay

The Analyst ◽  
2020 ◽  
Vol 145 (18) ◽  
pp. 6054-6060
Author(s):  
Juan Hu ◽  
Wen-can Li ◽  
Jian-Ge Qiu ◽  
BingHua Jiang ◽  
Chun-yang Zhang
Keyword(s):  
Energy Transfer ◽  
Fluorescent Probe ◽  
Fluorescence Resonance Energy Transfer ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Nuclease Activity ◽  
Activity Assay ◽  
Dna Nanostructure ◽  
Ratiometric Detection ◽  
Fluorescence Resonance

We develop a four-color fluorescent probe for ratiometric detection of multiple nucleases based on multistep fluorescence resonance energy transfer.

A quantum-dot based protein module for in vivo monitoring of protease activity through fluorescence resonance energy transfer

2011 ◽  
Vol 47 (18) ◽  
pp. 5259 ◽  
Author(s):  
Payal Biswas ◽  
Lakshmi N. Cella ◽  
Seung Hyun Kang ◽  
Ashok Mulchandani ◽  
Marylynn V. Yates ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Quantum Dot ◽  
Fluorescence Resonance Energy Transfer ◽  
Protease Activity ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
In Vivo Monitoring ◽  
Fluorescence Resonance
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