super-resolution imaging
Better in 3D Nanomachines observed in living cells Through a combination of genetic engineering, super-resolution microscopy and biocomputation, European researchers have been able to observe protein nanomachines in living cells in 3D.
A
On the left, an in vivo image of nanomachines using current microscopy techniques; on the right, the new method allows 3D observation of nanomachines in vivo and provides a 25-fold improvement in resolution. Image credit: O Gallego, IRB Barcelona.
lso known as protein complexes,
protein nanomachines are the structures responsible
angle from which the immobilised nanomachinery was viewed.
Gallego has already used the method to study exocytosis, a mechanism that the cell uses to
for performing cell functions. Currently, biologists
Then, in order to determine the 3D structure
communicate with the cell exterior. He and his
who study the function of these complexes isolate
of the protein complex, they used super-resolution
colleagues have now revealed the entire structure
them in test tubes, divorced from the cell, and
techniques to measure the distances between
of a key nanomachine in exocytosis that was until
then apply in vitro techniques that allow them to
different components and then integrate them in
now an enigma.
observe their structure up to the atomic level. They
a process similar to that used by GPS.
“We now know how this machinery, which
can also analyse these complexes within the living
The method enabled the researchers to observe
is formed by eight proteins, works and what each
cell, but that provides little structural information.
protein complexes with a precision of 5 nm — a
protein is important for,” said Gallego. “This
“The in vitro techniques available are excellent
resolution four times better than that offered by
knowledge will help us to better understand the
and allow us to make observations at the atomic
super-resolution, according to Gallego. By directly
involvement of exocytosis in cancer and metastasis
level, but the information provided is limited,”
observing the structure of the protein machinery
— processes in which this nanomachinery is altered.”
said Oriol Gallego, who coordinated the study
in living cells while it is executing its function, the
Furthermore, an understanding of how
group. “We will not know how an engine works
scientists have enabled cell biology studies that
nanomachines carry out their cell functions has
if we dissemble it and only look at the individual
were previously unfeasible.
biomedical implications, since alterations in their
parts. We need to see the engine assembled in the
inner workings can lead to the development of
car and running.”
diseases. The new strategy could therefore make it
Seeking a solution, Gallego and his colleagues
possible to see how viruses and bacteria use protein
at IRB Barcelona, in collaboration with the
nanomachines during infection, as well as better
University of Geneva and the Centro Andaluz de
understand the defects in complexes that lead to
Biología del Desarrollo, developed a strategy that
diseases in the first place.
brings together methods from super-resolution
Gallego said the ability to see 5 nm protein
microscopy, cell engineering and computational
complexes is “a great achievement”, though he
modelling. Their work was published in the
added that there is “still a long way to go to be able
journal Cell. The scientists began by genetically modifying cells in order to build artificial supports inside onto which they could anchor protein complexes. These supports would allow the researchers to regulate the
Model of the architecture of the main machinery involved in exocytosis. The eight proteins, each shown in a different colour, are bound, forming the nanomachine. Image credit: O Gallego, IRB Barcelona.
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to observe the inside of the cell at the atomic scale that in vitro techniques would allow”. “I think that the future lies in integrating various methods and combining the power of each one,” he said.
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