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From the structure and function to the design of modular proteins

Objectif



Protein modules constitute a distinct subset of protein domains, identified by sequence motifs. They are used repeatedly as "building blocks in functionally diverse proteins. In the course of evolution, nature has used protein modules as a way to generate rapidly and efficiently new proteins to respond to specific functional needs. Module have provided structural elements that have been reshuffled and reused sometimes over thousands of times. Some have been adapted to give specific interactions while others appear to have remained structural elements. At present, over 60 protein modules have been identified. They are used mostly in extracellular proteins of eucaryotes. The three dimensional structure of only a few modules has been determined. The objective of our project is to increase our understanding of the structure/function relationships of modular proteins, with the aim of designing novel modular proteins of biomedical and/or industrial value.
The focus of the research project will be on a limited number of modules that are particularly well suited to decipher the "language" of modules. Six types of modules have been chosen. Ig, FN3, and EGF modules are used ubiquitously as structural building blocks and in many interactions. C-type lectin, Clq, and Gla modules are more specialised and promote specific interactions with other biomolecules, glycans, antibodies and phospholipid membranes, respectively. Since many functionally important interactions are weak and require clustering of modules, the process of their oligomerization will also be analysed. The research project has three levels. At the first level, we will determine the 3D structure of individual modules and the structure and dynamics of arrays of modules, mostly produced by genetic engineering. By analysing the interactions of adjacent and reshuffled modules we will establish the rules that govern the shapes and the dynamic plasticity of modular proteins. This part of the study will require the use and the development of state of the art techniques for protein dynamics investigation, coupled with use of computer modelling techniques.
A second level of study will be to determine how modules interact. This will be achieved using a combination of surface plasmon resonance analysis and site-directed mutagenesis. This information integrated with structural and dynamic data will provide a detailed functional map of interacting module surfaces.
At the third level of the project, the knowledge gained by all participants on the structure, organization and function of modules will be integrated in the effective design of novel proteins. Three prototype modular proteins (modulins) with predefined properties are planned to test the predictions. Modulin 1 will have pre-set structural properties.
In modulin 2, modules will be clustered through oligomerization to increase the binding capacity of the final molecule. Modulin 3 Will include a module targeted to a chosen tissue and additional modules able to trigger specific cell responses.

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CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
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