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Using LANL GFP for Drug DiscoveryOverviewAnother valuable use of LANL's GFP technology is to determine the effects of other molecules (small molecules, antibodies, aptamers, etc.) on a specific protein of interest. Determining such an effect could be done a couple of different ways depending upon which GFP technology will be employed. For example, if a researcher wants to study a protein that is naturally insoluble due to genetic mutation, that protein could be transformed into E. coli and expressed in a way similar to that described on the Solubility page of this website, which describes a split version of GFP. However, instead of immediately expressing the S1-10 tag to complement the GFP tether, one could treat the cell with a library of small molecules first, then express S1-10. If a protein flouresces after treatment of a particular small molecule, whereas it did not before the treatment, it is safe to assume that small molecule has helped to correct the protein's solubility in some way. In a related way, our Folding Reporter could also be used to determine effects of other molecules on a protein of interest. The Folding Reporter reports on the success of a target protein’s folding and solubility. When LANL's engineered version of GFP is attached to the C-Terminus of a target protein, it mimics the folding success of that target protein. Thus, when the target protein folds correctly, GFP folds correctly and fluoresces. Conversely, when the target protein folds incorrectly, GFP folds incorrectly and fails to fluoresce. The Folding Reporter mimics the folding state of the target protein and the level of fluorescence is correlated with how well the target protein is folded. At LANL, Folding Reporter’s key application is evolving proteins for solubility in a high throughput format. But it could easily be used to determine the effect a particular cell treatment has on the target protein in a fast, high-throughput way. A researcher would simply need to place cells in a 96- or even 384-well format, treat each well with a different compound and quickly determine improved flourescence. In fact, this approach has been proven successful by Michael Hecht's group at Princeton doing research on Alzheimer's (see the technical library for more information). It is also being pursued by Dr. Greg Philipps at the University of Iowa to investigate RNA-protein interactions in vivo in E. coli (read the user feedback page). Please see our technical library for a long list of citations related to our Folding Reporter. Key Characteristics
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