First 3D Observation of Nanomachines Working Inside Cells
Researchers headed by IRB Barcelona combine genetic engineering, super-resolution microscopy and biocomputation to allow them to see in 3D the protein machinery inside living cells
Published in the journal Cell, the study unveils key functional features of an assembly of proteins that is vital for animals and plants.
Today scientists at the Institute for Research in Biomedicine (IRB Barcelona) present a study in Cell where they have been able to observe protein nanomachines (also called protein complexes)—the structures responsible for performing cell functions—for the first time in living cells and in 3D. This work has been done in collaboration with researchers at the University of Geneva in Switzerland and the Centro Andaluz de Biología del Desarrollo in Seville.
“The in vitro techniques available are excellent and allow us to make observations at the atomic level, but the information provided is limited. We will not know how an engine works if we dissemble it and only look at the individual parts. We need to see the engine assembled in the car and running. In biology, we still do not have the tools to observe the inner workings of a living cell, but the technique that we have developed is a step in the right direction and we can now see, in 3D, how the protein complexes carry out their functions,” explains Oriol Gallego, IRB Barcelona researcher and coordinator of the group that undertook this study, which also involved PhD student Irene Pazos.
Watching the nanometric machinery at work
The new strategy brings together methods from super-resolution microscopy—a discovery that was recognised with the 2014 Nobel Prize in Chemistry—, cell engineering, and computational modelling. The technology allows us to observe protein complexes with a precision of 5 nm*, a resolution “four times better than that offered by super-resolution and that allows us to perform cell biology studies that were previously unfeasible,” explains Gallego. (*a nm is a millionth part of a mm. A hair has a width of 100,000 nm)
Basic features of exocytosis
Gallego has used this method to study exocytosis, a mechanism that the cell uses to communicate with the cell exterior. For instance, neurons communicate with each other by releasing neurotransmitters via exocytosis. The study has allowed the scientists to reveal the entire structure of a key nanomachine in exocytosis and that until now was an enigma. “We now know how this machinery, which is formed by eight proteins, works and what each protein is important for. This knowledge will help us to better understand the involvement of exocytosis in cancer and metastasis—processes in which this nanomachinery is altered,” he explains.
New studies
An understanding of how nanomachines carry out their cell functions has biomedical implications since alterations in the inner workings can lead to the development of diseases. With this new strategy in hand, it will be possible to study cellular protein machinery in health and in disease. For example, it would be possible to see how viruses and bacteria use protein nanomachines during infection, and to better understand the defects in complexes that lead to diseases in order to design new therapeutic strategies that reverse them.
Over five years, Oriol Gallego has developed this project in the Molecular Medicine Programme at IRB Barcelona through a Ramón y Cajal researcher contract awarded by the Ministry of Economy and Competitiveness and that will be ending soon. Gallego has already lined up two research placements, in Japan and Germany, to learn more about integrating microscopy techniques. “After, I would like to continue to do top-level research in Barcelona, and I hope that this study that has been published in Cell helps me to do so,” comments the young researcher, whose focus lies in protein complex biology and in developing the technology that “makes the invisible visible”.
Contacts and sources:
Institute for Research in Biomedicine-IRB
Citation: The in vivo architecture of the exocyst provides structural basis for exocytosis. Authors: Andrea Picco, Ibai Irastorza-Azcarate, Tanja Specht, Dominik Böke, Irene Pazos, Anne-Sophie Rivier-Cordey, Damien P. Devos, Marko Kaksonen, Oriol Gallego Cell (2017). Doi: 10.1016/j.cell.2017.01.004
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