Abstract:
This research investigates the efficacy of adenosine analogs in inhibiting Chikungunya virus (CHIKV) replication by targeting the S-adenosyl methionine-dependent methyltransferase (MTase) activity of non-structural protein 1 (nsP1). Despite the health threat posed by alphaviruses such as CHIKV, effective antivirals remain limited. Previous studies have lacked direct, non-radioactive methods for measuring nsP1 MTase activity. Addressing this gap, we aimed to develop a capillary electrophoresis (CE)-based assay to screen potential MTase inhibitors efficiently. Methodologically, the study involved cloning, expression, and purification of nsP1 from both CHIKV and Venezuelan equine encephalitis virus (VEEV). This CE assay enabled direct detection of MTase reaction products, facilitating reliable screening of adenosine analogs. Results highlighted 5-iodotubercidin (5-IT) as a potent inhibitor, significantly reducing CHIKV replication in Vero cells with an EC50 of 0.409 μM. The main takeaway is that 5-IT, identified through a robust CE-based assay, offers promise as an antiviral candidate targeting alphaviral MTase activity.
Conclusion on the Role of Constant Systems Cell Disruption equipment:
The Constant Systems Equipment, specifically the French press, was instrumental in this research by enabling efficient and high-quality cell lysis for the extraction of nsP1 proteins. This step was critical to obtaining the purified protein necessary for assay development and reliable MTase activity measurements, underscoring the role of effective equipment in molecular studies targeting viral replication processes.
