Friday 3 April 2015

First sensor for 'crowd control' in cells

Scientists have developed a molecular sensor to measure 'crowding' in cells, which reflects the concentration of macromolecules present. The sensor provides quantitative information on the concentration of macromolecules in bacteria and in mammalian cells.

Living cells are full of macromolecules like proteins and nucleic acids. This has a profound influence on the way molecules inside a cell interact. Crowding reduces diffusion, but it also means that molecules stay together more easily. For example, DNA transcription is a hundred times faster under realistic cellular conditions than under diluted laboratory conditions. Despite its importance, crowding is rarely taken into account in biochemical studies.

The main reason for this is the lack of a reliable measurement technique for crowding. So far, it was only possible to estimate crowding from the average concentration of macromolecules and average cellular volumes. The new sensor is able to measure crowding in living cells, with a resolution that allows for the visualization of intracellular differences in both time and space.

The new sensor was developed by Dr Arnold Boersma and Professor of Biochemistry Bert Poolman. They designed a protein 'spring' with fluorescent protein markers on both ends. The first marker emits a blue light when excited by laser light. This blue light in turn excites the second marker, which then emits yellow light. This transfer of resonance energy is proportional to the distance between both markers; the technique is called 'Förster resonance energy transfer' (FRET).

Macromolecules exert a mechanical pressure on the protein spring, forcing the markers closer together. A series of control experiments have ruled out that other forces (e.g. ionic strength or chemical affinity) affect the distance between both markers. Other experiments have shown that the sensor gives an accurate quantitative estimate of crowding in cells.

For further details see:

Arnold J Boersma, Inge S Zuhorn, Bert Poolman. A sensor for quantification of macromolecular crowding in living cells. Nature Methods, 2015; DOI:10.1038/nmeth.3257

Posted by Tim Sandle

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