UROP Proceedings 2022-23

School of Science Division of Life Science 15 Cryo-EM Study of Membrane Proteins Supervisor: DANG, Shangyu / LIFS Student: TEMIRBEKOV, Yerassyl / SSCI Course: UROP1100, Summer TMEM168 is an evolutionary conserved eukaryotic membrane protein with an unknown structure and unclear function, appearing in a diverse set of research contexts, that has been shown to have many loosely related interactions. This protein has no apparent homologs outside the TMEM168 family; the sequence analysis shows no defined known domains, which may indicate novel functional mechanisms. We aim to overexpress and purify the target protein and later use it to resolve structure using single-particle cryo-EM and unravel functional role using biochemical methods. Understanding the structure and function of TMEM168 can lead to the discovery of new signal pathways, regulatory mechanisms, and other previously unknown molecular interactions. Analysis of Export of a Planar Cell Polarity Protein, Vangl2, out of the Trans Golgi Network Supervisor: GUO, Yusong / LIFS Student: CHENG, Guo / BCB-IRE Course: UROP1100, Fall UROP2100, Spring In intracellular trafficking, Trans-Golgi Network (TGN) is the transportation hub that sorts proteins to various downstream destinations including endosomes, plasma membrane, etc. The surface delivery of Vangl2, a conserved signaling receptor regulating Planar Cell Polarity (PCP), has been one important research direction in understanding the elaborate sorting mechanisms. It was previously demonstrated that both AP1 and Arfrp1 are involved in the transportation of Vangl2 from TGN to the plasma membrane. (Guo et al., 2013) Here we purified AP1 core, Arfrp1 and HAVangl2 to study the interaction between AP1 core and Vangl2, as well as the function of Arfrp1 in regulating the TGN export of Vangl2. Analysis of Surface Delivery of Epidermal Growth Factor Receptors Supervisor: GUO, Yusong / LIFS Student: CHU, Man Kit Christopher / BCB Course: UROP1000, Summer Epidermal Growth Factor Receptor (EGFR) is a well-known biomarker for cancer studies. Newly synthesized EGFR follows conventional protein trafficking route, it first transports from the endoplasmic reticulum (ER) to the Golgi and then delivered to the plasma membrane, allowing ligand binding to activate the EGFR signaling. This project aims to identify a nanobody that can block the export of EGFR and causes its retention at the ER membrane, thereby reducing the abundancy of EGFR on the cell surface to mitigate the activation of EGFR signaling. To commence, we first generated two different “Twin-Strep-EGFR” constructs through molecular cloning. These constructs are used for protein purification and later used for screening nanobody that targets EGFR.