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New probes from Scripps research quantify folded and misfolded protein levels in cells
Date:3/4/2014

udy, however, the researchers used folding probes in combination with cell lysis and ATP depletion, which causes the chaperones in the cell to hold onto the unfolded proteomepreventing its folding, providing a snapshot of the folded protein-of-interest population, while minimizing overrepresentation of that state by the tagging process.

One of the most important applications of new probes like these will be for the rapid, "high-throughput" screening of very large drug compound libraries to identify drug candidates that prevent protein misfolding by improving the quality of cellular folding. "Using these probes to quantify the concentration of a functional, folded protein-of-interest, we can screen for compounds that boost this concentration, for example," said Zhang, who along with Kelly conceived and designed the study.

In this study, the researchers cleared another hurdle for the use of probes in high-throughput screens with the design of a probe whose fluorescent beacon isn't lit all the time, but only turns on when it reacts with the folded protein-of-interest. "That fluorescent signal quickly shows you the concentration of the folded, functional protein that was in the cell at the time of lysis," Zhang said. "There is no need for the time-consuming removal of fluorescence probes that aren't bound to targets or separation of the probeprotein-of-interest conjugate."

Drugs that reduce the misfolding of specific proteins by altering cell biology of protein homeostasis might one day be used to prevent or delay age-related neurodegenerative diseases, such as Alzheimer's and Parkinson's, and to treat inherited enzyme-deficiency disorders. Zhang, Kelly and their colleagues suspect that anti-misfolding drugs could have even broader applications, given the surprisingly large populations of misfolded proteins in cells and all the ways in which those misfolded proteins could be causing harm.


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Contact: Mika Ono
mikaono@scripps.edu
858-784-2052
Scripps Research Institute
Source:Eurekalert  

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