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Contenido archivado el 2024-06-10

The nature of dynamic protein - protein interactions

Objetivo



Research objectives and content The alm of the research is to elucidate the nature of dynamic protein:protein interactions. Why are some complexes of proteins extremely static while others, in particular complexes of redox proteins, have very high turnover rates ? What are the roles in complex formation of the various non-covalent forces, such as electrostatic and hydrophobic forces and hydrogen bonds ? The project will analyze the structural and dynamic properties of a dynamic protein:protein complex in detail by a combination of kinetic measurements, site-directed mutagenesis and a new NMR approach. The latter is based on analysis of chemical shift changes caused by a) binding effects and b) intermolecular paramagnetic effects. This method is an excellent tool to obtain detailed information on the interaction sites of the two proteins and it has made it possible to determine the structure the complex of plastocyanin ( 10 kDa) and cytochrome (28 kDa), two redox proteins that have a crucial function in photosynthesis. In the project this new method will be exploited to study the effects on complex formation of the various non-covalent forces separately. Mutagenesis will be used to manipulate properties such as charge distribution and surface shape and polarity and the complexes of these mutants proteins will analyzed with NMR and by kinetic measurements. Furthermore, NMR will be used to study the dynamic properties of the proteins in the complex. Training content (objective, benefit and expected impact) Training objectives are twofold. First, to become fully competent as an experimental NMR spectroscopist, able to use multidimensional techniques for the study of proteins, and, second, to get familiar with the latest-developments in methods for site-directed mutagenesis and protein purification.

Ámbito científico (EuroSciVoc)

Programa(s)

Tema(s)

Convocatoria de propuestas

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Régimen de financiación

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Coordinador

Aristotle University of Thessaloniki
Aportación de la UE
Sin datos
Dirección
Analytical Chemistry Lab
54006 Thessaloniki
Grecia
Coste total
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