“When in doubt bind DFG-out”
Eric graduated from Saint John Fisher College in 2011 with a B.S. in Chemistry w/ a concentration in Biochemistry. While an undergraduate Eric studied the utility of electrophilic reactions of arene oxide-oxepin systems under Dr. Daniel Piccolo. In 2012, he joined the lab of Dr. Matthew Soellner in the Medicinal Chemistry department at the University of Michigan. In the Soellner Lab, Eric has performed phenotypic screens for target and lead compound discovery in understudied pediatric cancers using a profiled kinase inhibitor library. During a 10-week sabbatical, he validated the findings of this screen in Synovial Sarcoma in the lab of Dr. Torsten Neilsen at the University of British Colombia. Additionally, Eric has developed novel selective fluorescent probes to visualize subcellular localization of kinases in live-cell confocal microscopy. He is a recipient of the American Foundation for Pharmaceutical Education and Chemistry Biology Interface Training Program grants.
When not in the cell or chemistry hoods, Eric enjoys competing in basketball and football, critically evaluating films, and cheering on Michigan Sports (Go Blue!).
Development and use of a c-Src selective small molecule turn-on fluorescent probe
The subcellular localization of kinases is an important regulator of their activation and function. Common techniques used in fluorescent imaging of kinase localization, such as genetic fluorescent tags (i.e. GFP) and immunofluorescence, have proven useful; however, each has their own inherent disadvantages. Genetic fluorescent tags alter expression levels which can affect localization, and the fluorescent protein itself can affect function of target. Additionally, this strategy is only possible in cell models amenable to genetic manipulation. Immunofluorescence, which requires fixing (i.e. killing) of the cells, fails to capture the dynamic nature of kinases and can produce artifacts. Small molecule fluorescent probes for kinases would offer complimentary advantages to these methods.
We developed a highly selective and versatile fluorescent probe for c-Src, a non-receptor tyrosine kinase and a key signaling component in multiple malignancies. Our probe, which combines the kinase binding head group PP2 and a Coumarin fluorophore, covalently binds through the non-conserved C280 of c-Src through an electrophilic moiety within the linker. This PP2-Coumarin probe displays turn-on fluorescence, can be used in live-cell microscopy for the subcellular visualization of endogenous c-Src, does not require washing, and is compatible with super-resolution live-cell STED microscopy. We also demonstrate that this strategy can be applied generally to other kinases of interests. We have since used PP2-Coumarin to profile differential c-Src localization within a panel of Triple Negative Breast Cancer cell lines. Using this probe, preliminary results have shown that conformation selective inhibitors influence c-Src localization through the binding of active and inactive conformations of c-Src.