Semester of Graduation
Master of Science (MS)
Notch proteins are transmembrane proteins and act as receptors in Notch signaling, which is related to diverse developmental diseases including cancer.1-2 The mouse Notch1 protein contains 36 tandem epidermal growth factor-like (EGF) repeats in the extracellular domain (ECD).1-2 Our research project focused on the structural study of the Abruptex region consisting of EGF24-29 in the ECD. The fact that this region is a potential competitor to a Notch ligand, because of its high glycosylation and flexibility, makes it important to solve the three-dimensional structure and investigate the glycosylation effect on the structure of the Abruptex region.3 By doing so, we can have a better understanding of the binding mechanism of Notch receptors and the ligands. EGF27 from this region was selected as our first target protein domain because it is the only EGF repeat in this region that contains both O-glucosylation and O-fucosylation sites.1 In order to obtain NMR spectra of isotopic labelled EGF27 with high resolution, we first used the enhanced green fluorescent protein (EGFP) as a fusion of EGF27 as mentioned in Chapter 2. More sequences and E. coli strains were tested for a better yield of the target protein in Chapter 3, whereas Chapter 4 mostly describes the progress in the purification procedure and the characterization of EGF27 using nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS).
Zhong, Huimin, "Expression, Purification and Characterization of EGF27 from the Notch1 Abruptex Region" (2017). LSU Master's Theses. 4352.
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