Fluorescence spectroscopy method charts protein networks in living cells

An interdisciplinary team of researchers from the Cell Biology and Biophysics Unit, the Ellenberg Laboratory, and the Advanced Light Microscopy Facility at the European Molecular Biology Laboratory (EMBL; Heidelberg, Germany) has developed a fluorescence spectroscopy-based method to study the behavior of proteins in living cells. The approach allows scientists—for the first time—to follow the protein networks that drive a biological process in real time.

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Which proteins interact with each other and where they meet within cells is of huge interest to scientists because it reveals the state and activity of the molecular machinery that drives the most fundamental functions of life, such as the ability of cells to divide. The new technique will also be useful for scientists to investigate the mechanisms of disease and for pharmaceutical companies to explore new drug targets.

At the heart of this technology is a method called fluorescence correlation spectroscopy (FCS). Originally developed in the 1970s, FCS enables scientists to track individual proteins inside living cells once they have been fused to a fluorescent marker. FCS can measure where proteins move and when they meet, in a similar way to how Google can track people in traffic jams with their smartphone GPS signals.

Although such data would be tremendously valuable to have for all the proteins within a cell, until now FCS microscopes have been very cumbersome to use and the data difficult to interpret. The new development at EMBL automates the whole process of measuring protein behavior and analyzing the very large amounts of single-molecule data.

"Because we have made FCS into a high-throughput method, we can acquire data from many different proteins, which is key to studying biological networks that typically consist of tens to hundreds of components," says Jan Ellenberg, Head of Unit and Senior Scientist at EMBL.

The research was jointly funded by EMBL and the European Union. The research team plans to exploit the method now to build up a "Google Map" of the proteins in a living cell.

Full details of the work appear in the journal Nature Biotechnology; for more information, please visit http://dx.doi.org/10.1038/nbt.3146.

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