Incorporation of EGFR and RON Receptors into Nanodiscs

Author

Cristina Flores-Cadengo, University of New MexicoFollow

Publication Date

Spring 4-12-2019

Abstract

Understanding the structure-function relationship of membrane receptors is essential to comprehend the crosstalk between key signaling pathways. Aberrant trans-activation between receptors can lead to tumorigenesis. Two of these receptors known to be involved in cancer development are receptor tyrosine kinases (RTKs), RON (Recepteur d'Origine Nantais) and EGFR (Epidermal Growth Factor Receptor). There has been evidence of heterodimerization and crosstalk between these two receptors based on co-immunoprecipitation, however the structural requirements behind these interactions remain unknown. Structural studies could provide insights into these RTKs’ modes of dimerization and structure-function relationship. However, structural studies of full-length membrane proteins are often difficult due to poor solubility of the hydrophobic transmembrane domains. This affects protein structure and functionality. The use of nanodiscs for protein structural studies helps provide a native-like environment for membrane proteins, helping to avoid denaturing and aggregation, as well as providing a homogeneous size which makes them ideal for imaging techniques. In this work, we focus on optimizing a nanodisc assembly protocol to incorporate full-length RON and EGFR receptors into nanodiscs, as well as developing techniques to detect protein incorporation into these nanodiscs, which would ultimately facilitate structural studies for RON and EGFR heterodimerization. These studies could provide a mechanistic justification for novel targeted therapies in cancer.

Language

English

Keywords

Nanodiscs, Receptor Tyrosine Kinases, Electron Microscopy, Single-molecule imaging, protein structural studies, Size-exclusion chromatography

Document Type

Thesis

Degree Name

Biomedical Engineering

Level of Degree

Masters

Department Name

Biomedical Engineering

First Committee Member (Chair)

Diane S. Lidke

Second Committee Member

Mara P. Steinkamp

Third Committee Member

Eva Chi

Recommended Citation

Flores-Cadengo, Cristina. "Incorporation of EGFR and RON Receptors into Nanodiscs." (2019). https://digitalrepository.unm.edu/bme_etds/24