Date of Completion

Spring 5-1-2016

Project Advisor(s)

James Cole

University Scholar Major

Structural Biology and Biophysics

Disciplines

Biochemistry | Biophysics | Immunity | Immunopathology | Molecular Biology | Structural Biology

Abstract

The innate immune system is one of the first lines of defense against pathogens. RIG-I like receptors (RLRs) are a class of cytosolic receptors that recognize molecular patterns of invading pathogens and signal for downstream interferon induction. The RLR family consists of three proteins, RIG-I, MDA5, and LGP2. MDA5 and LGP2 work together to recognize and bind long viral dsRNA. LGP2 is thought to regulate MDA5 activation, but little is understood about this process. Characterization of the mechanism of action of these receptors requires careful biophysical and biochemical analyses. I have developed expression and purification of methods LGP2 and MDA5 in E. coil, by determining an appropriate cell line for expression and optimizing purification conditions. These methods will be utilized for production of LGP2 and MDA5 in large quantities necessary for biophysical and biochemical analyses. This work has provided the initial steps necessary for characterizing activation of LGP2 and MDA5 that will be beneficial for understanding innate immune processes. 2

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