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Content archived on 2024-05-30

A novel class of genetically encoded sensors of membrane protein function and structure

Periodic Report Summary - MEMSENSORS (A novel class of genetically encoded sensors of membrane protein function and structure)

The main scientific objective of the MEMSENSORS project was to test the hypothesis that fluorophore anisotropy can serve as a basis of a range of genetically encoded sensors of membrane protein function and structure. If our hypothesis is correct, anisotropy of the fluorophore in fluorescent proteins (FPs) could serve as a basis for creating a range of novel probes of cellular processes that involve membrane proteins. Since membrane proteins are both scientifically and economically important, but also notoriously difficult to study, our results could have large scientific and economic implications.

The specific scientific objectives of the project were:
1) to show that observed fluorescence changes in our prestin-based constructs are due to changes in fluorophore orientation;
2) to deduce information about the structure of our prestin-based constructs from fluorescence imaging data and
3) apply our microscopy methods to other proteins.

The scientific objectives were largely achieved, and in some cases substantially exceeded. Crucially, we have been able to show that fluorophore anisotropy can indeed be observed in a large number of membrane proteins in living cells, and that the data from our imaging experiments can be used to derive quantitative information about membrane protein structure and function. The imaging technique we have developed (two-photon polarisation microscopy) is applicable to a large number of proteins and questions, of high scientific and commercial relevance. Importantly, the technique holds promise in development of a usable voltage sensitive fluorescent protein, capable of detecting voltage pulses carrying information in living neurons. Such a protein would revolutionise the way much of neuroscience research is being done.

The project has produced and is still expected to produce a number of outcomes. A major result of the project is a device that allows precise monitoring of conformational changes in proteins, in living cells, on a sub-millisecond timescale. In developing the device, we collaborated with commercial entities. The device, as well as the associated microscopic method are the subject of a Czech patent that has now been awarded, as well as a PCT patent application that is being considered. Efforts to commercialise the device and method are underway. Apart from potential commercial outcomes, the project has also yielded significant scientific outcomes.

The results have been presented at several scientific meetings, and received exceedingly well. Several scientific publications in prestigious journals are expected in the coming months (one of our manuscripts is currently in second review in Nature Methods). Non-scientific objectives of the project (integration of the main researcher into the new country, furthering professional development, transfer of knowledge to host institution, cooperation with the non-associated third country) have largely been achieved.