Tiffany Cheng

Neurolysin is a mitochondrial zinc metallopeptidase involved in processing of a number of bioactive peptides and it functions to protect the brain from developing stroke. The result of stroke in brain is partly due to neurolysin’s inability to generate enough neuropeptides which act as targets for human disorders. The goal of this study is to co-crystallize neurolysin with a small chemical activator, His-Tyr, to understand how this molecule stimulates the enzymatic activity of neurolysin. It was hypothesized that His-Tyr is able to modulate enzymatic activity of human and mice neurloysin, where the molecule can bind and activate the enzyme but not inhibit or overlap with the substrate. A series of recombinant neurolysin constructs were generated by X-ray crystallography to understand, at molecular level, how His-Tyr interacts with neurolysin enzyme. Purified proteins were subjected to crystallization screening, and the initial hit condition was further optimized. Human and mouse crystals were generated using the hanging drop vapor diffusion method in the presence of a solution mixture. Crystal structure, including the molecular composition as well as bonding flexibility, of neurolysin was collected by X-ray diffraction. This research study will serve to promote a better understanding of how potent drug therapies can protect the brain from stroke and prevent early development of stroke injury.