Protein interactions direct cellular functions and their responses to pathogens and are important therapeutic targets. Scientists from the GSF Research Centre for Environment and Health have recently developed a method enabling simultaneous visualization of individual proteins and their interactions in living cells.
This is achieved by engineering the proteins to constantly emit red or blue fluorescent signals and to produce an additional yellow fluorescent signal upon interaction (see image below). Dr. Ruth Brack-Werner, Director of the GSF Institute of Molecular Virology (IMV) explains the decisive advantage of the new approach: “ In previous assays, signals were generated only by interacting proteins, whereas the individual partners remained undetected. However, the absence of signals could not be used to rule out protein interactions since the absence of one or both interaction partners would have the same effect. To overcome this problem Brack-Werner and her team developed the so-called extended bimolecular fluorescence complementation (exBiFC) which allows simultaneous monitoring of individual proteins and their interactions.
Dr. Ruth Brack-Werner, Institute of Molecular Virology of the GSF [300 dpi resolution for print] Photo: private.
Brack-Werner and her colleagues’ groundbreaking research work focusses on mechanisms that control replication of the human immunodeficiency virus (HIV), which causes AIDS. “HIV replication is based on the interaction of cellular proteins with viral proteins. Interactions involving viral regulatory factors have a direct impact on the amount of virus produced by the HIV host cell”, Brack-Werner explains. “Preventing HIV proteins from interacting with their crucial partners is a promising approach to developing novel therapies.” Therefore the GSF-scientists developed and validated exBiFC with the HIV Rev protein, which is an accelerator of HIV production. Various assays investigating Rev interactions in artificial settings indicate that the activity of Rev depends on the interaction of Rev molecules with each other and with cellular proteins. The latter include Exportin 1, which transports proteins from the nucleus to the cytoplasm and RISP, a modulator of HIV gene expression discovered by the Brack-Werner team in previous studies. Brack-Werner and her team demonstrated that exBIFC allows visualization of interactions of Rev with itself and with Exportin1 and RISP in living cells. In addition they were able to compare the strengths of the interactions of Rev with its partners by analysing the intensities of the signals in cell images.
ExBiFC has a wide range of potential appllications and represents an important tool for the elucidation of protein interaction networks and discovery of novel antiviral factors. Thus exBIFC has an enormous potential in the battle against leading global health problems such as infectious diseases and cancers.GSF - Forschungszentrum für Umwelt und Gesundheit, Germany
Heinz Joerg Haury | EurekAlert!
Closing in on advanced prostate cancer
13.12.2017 | Institute for Research in Biomedicine (IRB Barcelona)
Visualizing single molecules in whole cells with a new spin
13.12.2017 | Wyss Institute for Biologically Inspired Engineering at Harvard
MPQ scientists achieve long storage times for photonic quantum bits which break the lower bound for direct teleportation in a global quantum network.
Concerning the development of quantum memories for the realization of global quantum networks, scientists of the Quantum Dynamics Division led by Professor...
Researchers have developed a water cloaking concept based on electromagnetic forces that could eliminate an object's wake, greatly reducing its drag while...
Tiny pores at a cell's entryway act as miniature bouncers, letting in some electrically charged atoms--ions--but blocking others. Operating as exquisitely sensitive filters, these "ion channels" play a critical role in biological functions such as muscle contraction and the firing of brain cells.
To rapidly transport the right ions through the cell membrane, the tiny channels rely on a complex interplay between the ions and surrounding molecules,...
The miniaturization of the current technology of storage media is hindered by fundamental limits of quantum mechanics. A new approach consists in using so-called spin-crossover molecules as the smallest possible storage unit. Similar to normal hard drives, these special molecules can save information via their magnetic state. A research team from Kiel University has now managed to successfully place a new class of spin-crossover molecules onto a surface and to improve the molecule’s storage capacity. The storage density of conventional hard drives could therefore theoretically be increased by more than one hundred fold. The study has been published in the scientific journal Nano Letters.
Over the past few years, the building blocks of storage media have gotten ever smaller. But further miniaturization of the current technology is hindered by...
With innovative experiments, researchers at the Helmholtz-Zentrums Geesthacht and the Technical University Hamburg unravel why tiny metallic structures are extremely strong
Light-weight and simultaneously strong – porous metallic nanomaterials promise interesting applications as, for instance, for future aeroplanes with enhanced...
11.12.2017 | Event News
08.12.2017 | Event News
07.12.2017 | Event News
13.12.2017 | Health and Medicine
13.12.2017 | Physics and Astronomy
13.12.2017 | Life Sciences