PROSA · European Network on selection and analysis of protein-protein Interactions
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2006-03-01 → 2010-02-28
- EU contribution
- €2,368,446
- Participants
- 11
- Scheme
- RTN
Lines connect the coordinator with its partners. CORDIS does not always give exact coordinates for projects before 2014. These points are placed at city or country level.
Results in brief
Final Activity Report Summary - PROSA (European Network on selection and analysis of protein-protein Interactions)
Protein-protein interactions are key mediators of functions of life in the cell. They play crucial roles in regulation of enzyme action, in signal transduction and inter-cellular communication. The network developed new methodologies and approaches in which protein-protein interactions were studied, predicted, analysed or experimentally altered, probed or evolved. The SNAP-tag, commercialised by partner COVALYS and invented by partner EPFL was introduced as a new strategy for detecting protein-protein interactions in mammalian cell lysates based on the specific covalent cross-linking of two interacting partners. Computational analysis of proteins as network graphs provided a novel way for deriving functional predictions from structure and sequence data. Water-in-oil emulsion microdroplets were used to miniaturise biological assays to the pico-litre scale. Protein-protein interactions validated split-ubiquitin data were used to define cytokinesis events in the bud neck region of the S. cervisiae cells. In vitro evolutionary analysis gave insight on binding interaction involving protein-protein interfaces that share the evolvability features of enzymes, that is, the exploitation of promiscuous interactions and alternative binding configurations via generalist intermediates, and highlight the key role of compensatory stabilising mutations in facilitating the divergence of new functions.
Data: CORDIS, © European Union
Project objective
Networks of protein-protein interactions are central to cell and developmental biology and of great importance in molecular medicine. Given the potential importance of protein-protein interactions to healthcare, wealth creation and the biotech industry the re is an urgent need to train young researchers in the latest technologies aimed at identifying and dissecting such interactions by the very latest in high-throughput approaches, bio-informatics and rigorous biophysical analysis. We will train early-stage and experienced researchers to use new bio-informatics tools to direct high-throughput screening (HTS) for new protein-protein interactions. Network hubs" or "nodes" (i.e. proteins/protein families that interact crucially with one or many other partners in the network) will be identified and used as baits in HTS.Based on previous collaborative, published work, we will develop an in vitro selection system for protein-protein interactions by in vitro compartmentalisation (IVC). IVC relies on dispersing aqueous reactions into microscopic water-in-oil emulsion droplets ( ~5 fl) enabling selection of 10e10 pairs of interacting proteins. Hence, all ~10e9 possible pair wise binding interaction of proteins encoded by the human genome are possible in a more economical format than existing HTS techniques.A novel system for covalently labelling proteins will be used to label and cross-link proteins in vivo and to create protein micro-arrays. Kinetic and thermodynamic characterisation of identified protein-protein interact ions will be performed using a variety of techniques including a novel acoustic biosensor. Most importantly, we aim at an integrated approach where the bio-informatics tools we develop will be used to direct the HTS efforts. In turn, results obtained from t he screening will help validate and tune our newly-developed computational tools, while detailed mechanistic studies will provide insight into the physiological role of newly-identified interacting pairs."
Original text from CORDIS.
Participants
- THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIGE · CAMBRIDGECoordinatorCity levelUnited Kingdom
- COVALYS BIOSCIENCES AG · WITTERSWILCountry levelSwitzerland
- ECOLE POLYTECHNIQUE FÃÆ Ã Â©DÃÆ Ã Â©RALE DE LAUSANNE · LAUSANNESwitzerland
- FOUNDATION FOR RESEARCH AND TECHNOLOGY-HELLAS - FORTH · HERAKLION, CRETECountry levelGreece
- HELSINGIN YLIOPISTO · HELSINKIFinland
- MAX-PLANCK-GESELLSCHAFT ZUR FÃÆ Ã Â¶RDERUNG DER WISSENSCHAFTEN E.V. · MÃÆ Ã Â¼NCHENGermany
- UNIVERSITE LOUIS PASTEUR · STRASBOURGFrance
- UNIVERSITY OF YORK · YORKUnited Kingdom
- Universität Ulm · UlmGermany
- WEIZMANN INSTITUTE OF SCIENCE · REHOVOTIsrael
- Westfälische Wilhelms - Universität Münster · MünsterGermany
Links
Data: CORDIS, © European Union