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Method Article
A rapid assay for the biological evaluation of helicase activity.
Dimitrios Vlachakis
Bioinformatics and Medical Informatics Team, Biomedical Research Foundation, Academy of Athens
Corresponding Author
Sophia Kossida
Bioinformatics and Medical Informatics Team, Biomedical Research Foundation, Academy of Athens
Corresponding Author
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A new assay for the measurement of helicase enzyme activity was developed for the evaluation of the potency of potential inhibitors. This assay involves the use of a DNA or RNA duplex substrate and recombinant purified helicase. The DNA duplex consists of a pair of oligonucleotides, one of which is biotinylated and the other is digoxygenin (DIG)-labelled, both at their respective 5’ termini. This DNA duplex is immobilised, via the biotin molecule, on the surface of a neutravidin-coated 96 well plate. Helicase activity results in DNA unwinding upon activation by ATP, leading to the release of the DIG labelled oligonucleotides, which translates in signal (luminescence) reduction with respect to control wells. This signal can be produced and quantified with the aid of a chemiluminescence antibody.
Keywords
RNA, Helicase, Assay
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Subject Areas
Biochemistry
Biotechnology
Chemical biology
Structural biology
Microbiology
Biological techniques
Computational biology and bioinformatics
Associated Publications
Discovery of a novel HCV helicase inhibitor by a de novo drug design approach
by Sahar Kandil, Sonia Biondaro, Dimitrios Vlachakis, +6
Bioorganic & Medicinal Chemistry Letters (22 December, 2011)
Theoretical study of the Usutu virus helicase 3D structure, by means of computer-aided homology modelling
by Dimitrios Vlachakis
Theoretical Biology And Medical Modelling (22 December, 2011)
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This is a preprint. It has not completed peer review.
Method Article
A rapid assay for the biological evaluation of helicase activity.
Dimitrios Vlachakis
Bioinformatics and Medical Informatics Team, Biomedical Research Foundation, Academy of Athens
Corresponding Author
Sophia Kossida
Bioinformatics and Medical Informatics Team, Biomedical Research Foundation, Academy of Athens
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 22 Dec, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
A new assay for the measurement of helicase enzyme activity was developed for the evaluation of the potency of potential inhibitors. This assay involves the use of a DNA or RNA duplex substrate and recombinant purified helicase. The DNA duplex consists of a pair of oligonucleotides, one of which is biotinylated and the other is digoxygenin (DIG)-labelled, both at their respective 5’ termini. This DNA duplex is immobilised, via the biotin molecule, on the surface of a neutravidin-coated 96 well plate. Helicase activity results in DNA unwinding upon activation by ATP, leading to the release of the DIG labelled oligonucleotides, which translates in signal (luminescence) reduction with respect to control wells. This signal can be produced and quantified with the aid of a chemiluminescence antibody.
Figure 1
Figure 2
Figure 3
Associated Publications
Discovery of a novel HCV helicase inhibitor by a de novo drug design approach
by Sahar Kandil, Sonia Biondaro, Dimitrios Vlachakis, +6
Bioorganic & Medicinal Chemistry Letters (22 December, 2011)
Theoretical study of the Usutu virus helicase 3D structure, by means of computer-aided homology modelling
by Dimitrios Vlachakis
Theoretical Biology And Medical Modelling (22 December, 2011)