Penn Engineers have pioneered a brand new strategy to visualize the smallest protein clusters, skirting the physical limitations of light-powered microscopes and opening latest avenues for detecting the proteins implicated in diseases like Alzheimer’s and testing latest treatments.
In a paper in Cell Systems, Lukasz Bugaj, Assistant Professor in Bioengineering, describes the creation of CluMPS, or Clusters Magnified by Phase Separation, a molecular tool that prompts by forming conspicuous blobs within the presence of goal protein clusters as small as just a couple of nanometers. In essence, CluMPS functions like an on/off switch that responds to the presence of clusters of the protein it’s programmed to detect.
Normally, says Bugaj, detecting such clusters requires laborious techniques. “With CluMPS, you do not need anything beyond the usual lab microscope.” The tool fuses with the goal protein to form condensates orders of magnitude larger than the protein clusters themselves that resemble the colourful blobs in a lava lamp. “We expect the simplicity of the approach is certainly one of its foremost advantages,” says Bugaj. “You do not need specialized skills or equipment to quickly see whether there are small clusters in your cells.”
For treating diseases like Alzheimer’s, ALS and even cancer, with the ability to detect protein clusters this small guarantees to be a foundational advancement, allowing researchers to find out whether or not drugs actually eliminate disease-causing clusters of a goal protein in a cell.
“You wish a really clear signal,” says Bugaj, to know whether or not a treatment worked. “It’s extremely obvious when you have got a big cluster, but when you have got small clusters, it is far harder. Now we are able to amplify that signal and see which drugs actually dissolve the clusters.”
Along with providing latest avenues for drug discovery, CluMPS will permit researchers to grasp the functioning of proteins in latest ways, resulting in a deeper, more sophisticated rendering of cells themselves. “There’s a complete landscape of protein clustering that is happening on the small scale, that is essential, but we just don’t learn about it yet,” says Bugaj.
One among the challenges that CluMPS overcomes is that lightwaves themselves are larger than the smallest protein clusters, making it very hard to see such clusters without specialized techniques. “The wavelength of blue light is about 400 nanometers,” says Bugaj. “You possibly can’t actually resolve the placement of anything smaller than half that wavelength with a standard microscope,” rendering protein clusters tens of nanometers wide all but invisible.
To develop CluMPS, Bugaj and his lab partnered with Elizabeth Rhoades, Professor of Chemistry at Penn Arts & Sciences, whose lab helped validate that CluMPS did indeed detect goal protein clusters as an alternative of generating false positives. “It was a very rewarding collaboration for us,” says Rhoades, “since it allowed us to use the methods commonly utilized by our lab to assist validate this powerful latest tool in living cells. It was exciting to see how well we could differentiate between clusters and the only proteins.”
Thomas R. Mumford, a doctoral student within the Bugaj Lab and the paper’s lead writer, played a key role in brainstorming and performing the mandatory experiments. “It was crucial to characterize how underlying features of protein clusters interacted with CluMPS to trigger condensation,” says Mumford, to enable future users of the technology to grasp precisely how it really works. “The burden was on us to display that we were in reality detecting small clusters,” adds Bugaj. “One of the crucial rewarding points was working with Tom and the Rhoades lab to think about latest kinds of experiments that may convincingly make the purpose.”
