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Content archived on 2024-05-29

Development and application of time resolved resonance energy transfer (tr-FRET) based methods for investigation of the earliest stages of protein folding and misfolding

Objective

The question of how the sequence of amino acids dictates the final fold of globular proteins remains one of the fundamental riddles of structural biology. The objective of the present research plan is to identify sequence elements that contribute key local and non-local interactions that are effective during the initial steps of the folding process.

A series of experiments are proposed to identify critical residues that form such effective interactions. Initial phases of the misfolding of globular and natively unfolded proteins will be explored in order to explain these processes and ultimately to control negative consequences of misfolding, e.g. formation of amyloid plaques. An interdisciplinary approach aimed primarily at structural characterization of the rapidly changing ensembles of partially folded protein molecules will be adopted. A number of methods will be applied, in particular time resolved FRET, -FRET detected FCS; FRET detected fast-kinetic methods, (a novel high pressure jump and micromixing based double kinetics experiments); protein engineering and computational methods.

The hypothesis is that few very effective non-local contacts can direct the backbone into a native like fold at the early phase of the folding transition. This hypothesis will be explored to identify the essential key interactions and specific clusters of residues that contribute to those interactions. The response of the ensembles of conformers to changes of pressure; temperature; solvent composition and to the insertion of structure perturbation mutations will be monitored. Computational and bioinformatics methods will be applied in an attempt to generalize the experimental results.

In order to achieve these goals the host research group will bring in
- methods for construction of a pressure -jump and micromixing devices;
- methods of molecular dynamics simulations, and
- the technology enabling single molecule detection.

Fields of science (EuroSciVoc)

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Keywords

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Topic(s)

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Call for proposal

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FP6-2004-MOBILITY-3
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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.

TOK - Marie Curie actions-Transfer of Knowledge

Coordinator

BAR ILAN UNIVERSITY
EU contribution
No data
Address
Bar Ilan University Campus
RAMAT GAN
Israel

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Total cost

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.

No data

Participants (1)

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