MEASURING THE THERMODYNAMICS OF RECEPTOR TYROSINE KINASE INTERACTIONS: A STUDY OF HOMODIMERS, HETERODIMERS, AND RECEPTOR-ADAPTER PROTEIN COMPLEXES

thermodynamics;receptor tyrosine kinases;Materials Science & Engineering

Del Piccolo, NualaOstermeier, Marc ;

Johns Hopkins University

关键词: thermodynamics;    receptor tyrosine kinases;    Materials Science & Engineering;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/59321/dissertation_v3.docx?sequence=2&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

Protein-protein interactions regulate many biological systems. Understanding these interactions can provide insight into the function of proteins and the design of biological signaling networks, and can inform the development of more effective therapeutics. Protein-protein interactions can be described by thermodynamics, that is, the strength of binding, as well as by the distance between binding partners. In this thesis, I quantify protein-protein interactions in the Receptor Tyrosine Kinase (RTK) family, using novel quantitative fluorescence microscopy methods. In Chapter Three and Chapter Four, I characterize the formation of RTK homodimers using existing techniques. Specifically, I examine a subfamily of RTKs, known as the Fibroblast Growth Factor Receptors (FGFRs), in the presence of (i) pathogenic point mutations and (ii) activating ligands. In Chapter Five, I consider the formation of RTK heterodimers. I develop a method of measuring the thermodynamics of RTK heterodimer formation and apply it to the FGFR family. I also examine the effects of pathogenic point mutations on FGFR heterodimers. In Chapter Six, I study the interaction of RTKs with adapter proteins, an event that directly links RTKs to intracellular signaling networks. I develop and implement a procedure to measure the thermodynamics of this binding reaction, using the RTK Epidermal Growth Factor Receptor (EGFR) and the adapter protein Growth factor receptor-bound protein 2 (Grb2) as a model system. Overall, the work in this thesis demonstrates the utility of fluorescence microscopy for the quantitative characterization of protein-protein interactions in the membrane and reports previously unknown properties of the protein-protein interactions that regulate the RTK activation process.