Pictures of the formation of cells' "protein factories," has been revealed by the Scripps Research Institute's team, which will help in understanding the mysteries of the cell and disease treatments.
In addition to being a major technical feat on its own, the work could open new pathways for treatments for diseases tied to errors in ribosome formation.
In addition, the techniques developed in the study can now be applied to other complex challenges in the understanding of cellular processes.
Past studies of the intermediate molecules that combine to form ribosomes and other cellular components have been severely limited by imaging technologies.
Electron microscopy has for many years made it possible to create pictures of such tiny molecules, but this typically requires purification of the molecules.
To purify, you must first identify, meaning researchers had to infer what the intermediates were ahead of time rather than being able to watch the real process.
"My lab has been working on ribosome assembly intensively for about 15 years. The basic steps were mapped out 30 years ago. What nobody really understood was how it happens inside cells," said James Williamson, member of the Skaggs Institute for Chemical Biology.
The NRAMM group developed a new technique and dubbed discovery single-particle profiling, which dodges the purification problem by allowing successful imaging of unpurified samples.
An automated data capture and processing system of the team's design enables them to decipher an otherwise impossibly complex hodgepodge of data that results.
For this project, ribosome components were purified from cells of the common research bacterium Escherichia Coli.
Then chemically broke these apart to create a solution of the components that form ribosomes. The components were mixed together and then were rapidly stained and imaged using electron microscopy.
The team produced images that the scientists were able to match like puzzle pieces to parts of ribosomes, offering strong confirmation that they had indeed imaged and identified actual chemical intermediates in the path to ribosome production.
Interestingly, this work also confirmed that there are more than one possible paths in ribosome formation, a phenomenon known as parallel assembly that been suggested by prior research but never definitively confirmed.
The findings were published in the journal Science.