A New Target for Pain: Development of Tools to Study Human Histidine Triad Nucleotide Binding protein

The research described in the current thesis elucidates the contributions to the characterization of Hint1 from the aspect of function, mechanism, and structural determinants. To do so, we describe development of variety of tools to study Human Hint proteins. The first two chapters provide the first strong evidence of the role of Hint1 active site in the CNS using chemical genetics, medicinal chemistry and neuropharmacological studies. In Chapter 3, we describe the covalent capture of the Hint1-adenylated intermediate using an alternate substrate and time-lapse crystallographic studies as part of our efforts to elucidate reaction trajectory of Hint1. In Chapter 4, we describe the development of switch-on fluorescent probes as tools for monitoring the active site and detection of Hint proteins. Chapter 5 describes the structural and functional characterization of genetic mutations of Hint1 associated with peripheral neuropathy as well as structural determinants of Hint1.