Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1).
Nambisan, Anand Krishnan
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Disrupted in Schizophrenia 1 (DISC1) is a psychiatric disease risk gene implicated in numerous mental disorders including schizophrenia. Many isoforms of DISC1 are found in the human brain, of which the longest contains 854 residues. DISC1 is involved in all major stages of neurodevelopment; from proliferation of neural progenitor cells to neuronal migration and circuit integration. DISC1 functions like a scaffold, interacting with several protein partners, like Glycogen Synthase Kinase 3β (GSK3β) to dynamically coordinate different stages of brain development. GSK3β, a serine threonine kinase, plays an important role in the canonical Wnt/β-catenin signaling pathway. Mao et al. (2009) showed that DISC1 directly interacts and inhibits GSK3β in neural progenitor cells, thus promoting neurogenesis and proliferation. Our lab is interested in understanding the nature of this interaction and the residues involved in it. We followed two strategies to generate this DISC1-GSK3β complex and study its biochemical and biophysical characteristics. In the first method, we tried expressing and purifying the proteins individually and generating a stable binary complex in vitro. The second method involves the co-expression of DISC1 and GSK3β within the same cell, using a polycistronic vector, in attempts to form the interaction complex naturally. This thesis showcases the work done in optimizing the overexpression and purification protocols for obtaining pure DISC1 subdomains and as a stable complex illustrating its interaction with GSK3β as its protein partner. The thesis also focuses on understanding the biochemical and biophysical characterization of DISC1 independently as well as in its complex form.
DegreeMaster of Science (M.Sc.)
DepartmentVeterinary Biomedical Sciences
ProgramVeterinary Biomedical Sciences
CommitteeAdams, Gregg; Sanders, David; Dmitriev, Oleg
Copyright DateAugust 2020
Neuropsychiatric disorders Protein biochemistry
Protein expression and purification