Abstract 2010 Identifying the Oligomerization State of the Transmembrane Domains of Proteins Found in SARS-CoV-2

SARS-CoV-2, the virus responsible for the disease COVID-19, is a coronavirus containing multiple potentially targetable proteins.Some of these proteins are embedded in the SARS-CoV-2 viral envelope.The transmembrane domains of these proteins are highly conserved regions across all strains of coronaviruses.This research focused on the Membrane (M) and Open Reading Frame 3a (ORF 3a) proteins as potential oligomers to be studied for later research into targetable sites for inhibitors.The researchers chemically synthesized peptides corresponding to the transmembrane domain of each particular protein.The fluorophore nitrobenzodiazole (NBD) was then attached to the N-terminus of to one third of the peptide sample, and the fluorophore tetramethylrhodamine (TAMRA) was attached to the N-terminus of another third of the sample, while the last third of the sample was left unlabeled.Each of these groups was purified through high-performance liquid chromatography (HPLC) and analyzed by LC/MS.The oligomerization state was determined through a fluorescence assay using the fluorescently labeled peptides.Future work will further study the interactions in the transmembrane domain to determine if potential inhibitors could be designed to prevent function of the virus.