Objective
We propose a novel high-throughput method to study the chemo-mechanical coupling in molecular motors using helicases, enzymes that couple ATP hydrolysis to unwinding of the double helix, as a model system. The method combines a cell-free expression system, droplet microfluidics and fluorescent assays with next generation sequencing to test large (up to millions of elements) libraries of helicase mutants for both unwinding and ATPase activity. The aim is to identify all sequence positions that, if mutated, lead to uncoupling, i.e. abolish unwinding while preserving ATPase activity. Once these positions are identified, their distribution on the enzyme sequence and structure will be analysed to obtain insight into the coupling mechanism. Notably the methodology could be applied to systems where the classical, structure-based analysis of the chemo-mechanical coupling is not possible. We will demonstrate our method on the RecQ helicase from e. coli using existing information on this system as a means of validation. Any further insight will be a demonstration of the power of our approach. When fully developed, our method will serve as a discovery tool for the interdisciplinary community working on helicases that includes medical, biological and physical sciences. It may be applied to aspects of helicase biophysics beyond the chemo-mechanical coupling, to other enzyme families and to screen for helicase inhibitors with higher throughput and lower cost when compared to robotic mictotitre-plate techniques. The proposed work will be carried out at the Griffiths Lab (ESPCI Paris) with a secondment at the Crick Institute (London). This project integrates the training in biophysics of the Experienced Researcher with the expertise of the Host Institution on high-throughput methods and the experience of the Secondment Institution on helicases. The Experienced Researcher will be trained in next generation sequencing and fluorescence-based approaches to molecular motors.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences physical sciences classical mechanics fluid mechanics microfluidics
- natural sciences biological sciences biophysics
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
You need to log in or register to use this function
We are sorry... an unexpected error occurred during execution.
You need to be authenticated. Your session might have expired.
Thank you for your feedback. You will soon receive an email to confirm the submission. If you have selected to be notified about the reporting status, you will also be contacted when the reporting status will change.
Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
-
H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions
MAIN PROGRAMME
See all projects funded under this programme -
H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility
See all projects funded under this programme
Topic(s)
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Funding Scheme
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
MSCA-IF-EF-ST - Standard EF
See all projects funded under this funding scheme
Call for proposal
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
(opens in new window) H2020-MSCA-IF-2016
See all projects funded under this callCoordinator
Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.
75231 Paris
France
The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.