COSMO · Correlative smFRET/electrophysiology for long-term studies of protein dynamics

Correlative smFRET/electrophysiology for long-term studies of protein dynamics

Proteins carry out complex biological processes that are essential for cell survival. Rather than being static, they are highly dynamic molecules whose function depends on their 3D structure and dynamic properties. The study of protein dynamics is not only of fundamental interest, but also has direct implications for health and disease, as altered dynamics can lead to disease. However, it remains difficult to experimentally determine the multitude of conformations adopted by proteins, their lifetimes and the transitions between them. Single molecule FRET (smFRET) is probably the most suitable technique available but is limited by the short observation times (a few seconds) and the difficulty of measuring more than one molecular distance at the same time. The goal of this action “Correlative smFRET/electrophysiology for long-term studies of protein dynamics” is to develop an innovative correlative approach that can extend smFRET observations to much longer timescales, while also providing a way to integrate multiple measurements into a coherent molecular picture. How is it done? by combining smFRET measurements with nanopore-based electrophysiological experiments that have high temporal resolution and are also sensitive to conformational changes. Our correlative system thus combines the speed and sensitivity of electrical reading with the direct observation of molecular distances using smFRET. This new methodology should open entirely new opportunities to monitor the fast and long-term conformational dynamics of proteins or the assembly processes of macromolecules. This methodology is meant to map conformational dynamics of the biologically-important Ras proteins. Defects in Ras proteins are implicated in 30% of human cancers. We aim to better understand the conformational dynamics of selected Ras proteins with the ultimate goal of obtaining drugs that function through the regulation of these dynamics.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз