Allergies are an important health issue in our community. We want to develop a drug to lower the effects of allergies by characterizing the IgE receptor in a mast cell. The mast cell is an immune cell with many receptors on its membrane. Our project focuses on an IgE receptor called FcεR1, which plays a critical role in the allergy signal cascade within the mast cell.
The FcεR1 has three protein subunits: the alpha, beta, and gamma chain. In the gamma chain, there are two tyrosines (amino acid). The phosphorylation (transfer of a phosphate) of these tyrosines is known to be important for the initiation of signaling. However, the timeline and pattern of phosphorylation of each tyrosine is unknown. In our project, we will investigate these unidentified parameters using two independent methods: modeling and experimental biology.
These two approaches allow us to gain independent results that will complement and verify each other.
Using simulations, we calculated the rate of tyrosine phosphorylation. The experimental part of the project utilized viruses and yeast to select a specific antibody to the first phosphotyrosine.
This can detect when the first tyrosine is phosphorylated. An antibody specific to the second tyrosine is currently being developed.
We are currently investigating phosphorylation patterns of the gamma subunit.
Experimental results show that the first tyrosine is phosphorylated at two minutes and dephosphorylated by five minutes. Modeling simulations show phosphorylation at two minutes,
but no dephosphorylation by five minutes. We will also use modeling to investigate whether single or double phosphorylation is required to start the signal cascade. These results may eventually lead to finding small molecules that enhance/inhibit the signal cascade to develop a drug for allergies.
We want to develop drugs for allergies by characterizing an IgE receptor, FcεR1, in mast cells. Gamma chain phosphorylation in FcεR1 is important for mast cell signaling. We investigate patterns of gamma phosphorylation using modeling and experimental biology.
Simulations calculated rate of tyrosine phosphorylation. Experimentation selected an antibody that detects when the first phospho-tyrosine’s phosphorylation.
Experimental results show phosphorylation by two minutes and dephosphorylation by five minutes. Modeling shows similar results but without dephosphorylation by five minutes.
Modeling will investigate whether single or double phosphorylation initiates signaling.
Designing of small molecule drugs that block phospho-tyrosines will be based on results.