The SARS-CoV-2 virus, responsible for the ongoing global pandemic, has infected over 774 million people and caused more than 7.03 million deaths worldwide as of March 2024. Upon entering host cells, the virus’s positive-sense single-stranded RNA genome is translated into 29 viral proteins. Among the earliest expressed is a polyprotein that is cleaved into 16 nonstructural proteins (Nsps), including Nsp1, a key virulence factor that suppresses host immune responses by selectively inhibiting host protein translation. Cryo-EM studies have shown that residues 148–180 of Nsp1 adopt a helix-turn-helix motif upon binding to the 40S ribosome, effectively plugging the mRNA entry tunnel. At 1 μM, Nsp1 can reduce in vitro host translation efficiency by over 90%. The intrinsically disordered nature of its C-terminal (CT) domain presents challenges for structure-based drug design.

My research focuses on characterizing the structural dynamics and metal-binding properties of this disordered CT region. I have shown that aquo copper(II) binds selectively to histidine 165 in a synthetic 33-residue C-terminal peptide of Nsp1. Building on this, I am now examining how metal binding, including both copper(II) and cobalt(III) complexes, affects the structure and function of full-length Nsp1 and its variants. These studies aim to provide molecular-level insights into how transition metals may modulate Nsp1’s interaction with the host translation machinery.

Throughout this project, I have developed expertise in protein expression and purification, SDS-PAGE, mass spectrometry, and X-ray crystallography. I also use a suite of biophysical techniques, including tryptophan fluorescence (W161), time-resolved Förster resonance energy transfer (TR-FRET), electron paramagnetic resonance (EPR) spectroscopy, and circular dichroism. Complementing these tools, I perform in vitro translation assays to probe the functional impact of metal coordination on ribosome inhibition.