The caveolins are a family of human membrane proteins implicated in cancer and other diseases. In Chapter 1, I briefly explain some of the basic biochemistry of the caveolins, and why they are not only a promising target for research and therapeutics but also a particularly challenging target. The novel peptide WL47, the development of which is described in later chapters, is introduced as a caveolin-1 ligand. WL47 will allow new avenues of investigation for this challenging target.
In Chapter 2, I review, compare, and contrast some of the myriad techniques available to quantify binding affinity for high affinity binding interactions such as the interaction between caveolin-1 and WL47. As I discovered in my own attempts to measure the dissociation constant for this interaction, high affinity is a boon for future use of a ligand probe, but it can be challenging to quantify initially. Because a review of techniques specifically suited to high affinity measurement was not available at the time I needed it, I have written this chapter to fill this gap in the literature.
In Chapter 3, I discuss the development of peptide ligand WL47 through an iterative process of library design, synthesis, screening, and data analysis. High affinity binding, target selectivity, and activity as an inducer of deoligomerization of oligomeric caveolin-1 is presented. Beyond providing a novel tool for probing the function of caveolin-1 oligomerization, this chapter is also a roadmap for the creation of future peptide ligands through this generalizable, iterative process
Chapter 4 details the progress that has been made toward understanding the importance of WL47 dimerization through a disulfide bridge. This chapter also covers initial efforts towards designing a redox-stable version of WL47 that will remain functional in conditions like those found in the interior of a live cell.
Chapter 5 concludes the dissertation with a discussion of the current state of the caveolin ligand project, including detailed descriptions of the experiments that will be necessary for future researchers to bring this project to its conclusion.