To understand and modulate biological processes, we need their spatio-temporal molecular models. In view of the recent methodological and technical advances in fluorescence spectroscopy and microscopy as well as in multi-scale modeling of complex biochemical systems, the applicant proposed to build these models by a holistic approach. This goal corresponds to the development of a novel integrative platform for a Molecular Fluorescence Microscope (MFM) to achieve ultimate resolution in space (sub-nanometer) and time (picoseconds) for characterizing the structure and dynamics of proteins. The MFM will combine fluorescence spectroscopy with computer simulations in a hybrid approach, first, to derive a molecular description of all fluorescence properties of the tethered dyes in proteins (objectives 1 and 2) and, second, to utilize this information in simulations to report on the protein properties (objective 3). In this hybrid approach, high precision FRET measurements are the core experimental technique (hybridFRET). The MFM allowed us to tackle the central biophysical question of how intra- and intermolecular domain interactions modulate proteins' overall structure, dynamics, and thus ultimately function by spatio-temporal models (objective 4).
In 2015, no holistic use of fluorescence spectroscopy for structural modeling of proteins was reported. During the hybridFRET project, we established the proposed workflow to generate and deposit FRET-based structural models in the prototype archiving system for integrative structural models, PDB-Dev, so that the models are searchable and accessible for society.
