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Single protein folding Pathways

Objective

This project aims at importing in Europe key scientific competences developed by the applicant researcher during a period of almost 10 years spent at UC Berkeley (group of Prof. Carlos Bustamante) working on the technique of laser tweezers applied to the study of folding/unfolding pathways of single proteins.

The aforementioned group has pioneered the measurements of the tiny forces which allow a biopolymer to attain and maintain its native structure. Laser tweezers applied to protein folding, have the remarkable advantage of operating on a single molecule, avoiding the ensemble average typical of classical bulky techniques and providing naturally a reaction coordinate to follow the thermodynamic process of folding.

We propose to build the double trap laser tweezers at CNR-INFM-S3 in Modena and to investigate the energy landscape of the folding trajectories of the protein RNaseH by:
- estimating the activation energies between different molecular states along the reaction coordinate from the dependence of the folding rate constants on temperature;
- determining the structures of the transition states through Phi-value analysis;
- analyzing the anisotropy of the energy landscape by denaturing the molecules along different pulling axes, and
- carrying out molecular dynamic simulations to gain insight into the microscopic details of the folding reactions.

The results of these experiments should help elucidating the role of the energy landscape in determining the folding, dynamics and ultimately the function of a protein. The applicant researcher possesses all the needed capabilities to face this ambitious goal.

The host offers in this sense an ideally suited environment for the prompt transplantation and successful development of this technique in Europe. By this action we would import a very novel and unique skill that would foster further development in the field of single biomolecule investigation and nanobiotechnology.

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Coordinator

CONSIGLIO NAZIONALE DELLE RICERCHE

Address

Piazzale Aldo Moro 7
Roma

Italy

Administrative Contact

Paolo FACCI (Dr)

Project information

Grant agreement ID: 44952

  • Start date

    1 October 2006

  • End date

    30 September 2008

Funded under:

FP6-MOBILITY

Coordinated by:

CONSIGLIO NAZIONALE DELLE RICERCHE

Italy