Objective
Due to its intrinsic visible fluorescence, the Green Fluorescent Protein (GFP) is used to monitor cellular and biomolecular functions. Its chromophore is formed by cyclization of three aminoacid residues: Ser65, Tyr66 and Gly67. The substitution of Tyr66 b y Trp and few additional mutations lead to the Enhanced Cyan Fluorescent Protein (ECFP), characterized by cyan emission.
One unexplained feature is that ECFP has double humped rather than conventional single excitation and emission peaks. NMR and crystallo-graphic studies suggested that these two spectral peaks of ECFP originate from two conformational states of the protein. However, a preliminary time-resolved fluorescence study indicates that the photophysics of ECFP is more complicated. At least four lifetimes, which depend differently on pH and temperature are required to describe the fluorescence decay and suggest more than two fluorescent species.
Molecular calculations and simulations are particularly valuable for relating properties of biomolecules t o their structure. Our project will be the first study which will combine a variety of theoretical techniques and relate the results to spectral data, in order to understand the intriguing photophysics of ECFP. From electrostatic calculations we will obtain the population probabilities of the different states as a function of pH.
The analysis of molecular dynamics and of quantum mechanical calculations will provide information on the behaviour of each state and the temperature dependence of this behaviour. T he combination of these two approaches, together with the results of the excited state dynamics analysis, will lead to a quantitative mechanistic model of the dependence of the fluorescence signal on pH and temperature.
Understanding the structural basis o f the photophysical process will lead to the design of ECFP mutants with simplified or modified photophysical properties improving the fluorescence tools for cellular biology and medical diagnostics.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
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Keywords
Project’s keywords as indicated by the project coordinator. Not to be confused with the EuroSciVoc taxonomy (Fields of science)
Project’s keywords as indicated by the project coordinator. Not to be confused with the EuroSciVoc taxonomy (Fields of science)
Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Topic(s)
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
Call for proposal
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
FP6-2004-MOBILITY-5
See other projects for this call
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.
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.
Coordinator
BAYREUTH
Germany
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.