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RESEARCH

 

DESIGNING NEW FRET SENSORS TO UNDERSTAND THE INTRACELLULAR ENVIRONMENT

The interior of the cells is unlike dilute buffer. Yet, traditionally most biochemistry experiments have been performed in a dilute buffer. To understand how the interior of the cell influences biochemical processes, such as biocatalysis and organization of the biopolymers, we need to know what the important parameters in the cell are and quantify their contribution to the intracellular organization. 


By creative design, we developed a series of FRET sensors for macromolecular crowding and ionic strength: Cells are crowded with macromolecules that occupy space, generating a force that influences the organization of biomolecules in cells. The crowding sensors compress akin a polymer in crowded solutions. The sensors provide a readout for the excluded volume in bacteria, yeasts, and mammalian cells. Our efforts are towards improving these sensors, as well as applying them to different species. Further, we apply the sensors in dynamic soft materials.

Further reading:

Nature Methods, 2015, 227-229

Biophys J. 2017, 1929-1939

Nature Rev. Microbiol 2017, 309-318


Collaborators:

Prof. Veenhoff (UMCG Groningen)

Prof. Sheets and Prof. Heikal (Univ Minnesota)

Prof. Fitter (RWTH Aachen)

Dr. Aberg (UMCG Groningen)

Dr. Kedrov (HHU Düsseldorf)

IONIC STRENGTH SENSING

The other sensor that we developed measures the ionic strength. The ionic strength is important for anything charged, which includes pretty much every molecule in the living cell (apart from a few small molecules). Our FRET-based ionic strength sensor allows to determine the ionic strength inside living cells, with all the advantages of FRET-based genetically-encoded sensors, such as high spatiotemporal resolution, and easy targeting to different compartments.

Further reading:

ACS Chem Biol 2017, 2510-2514

LIFE-LIKE SYSTEMS, INFORMATION TRANSFER, DIAGNOSTIC TOOLS, AND MORE

We will provide more information in due course.

©2021 by Boersma Lab