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Abstract

Outbreaks of the Venezuelan Equine Encephalitis Virus (VEEV) and the Chikungunya Virus (CHIKV) continue to emerge in Central and South America, Africa, and Asia, but there are currently no vaccines or anti-virals for these viruses. Given their ease of transmission, debilitating symptoms, and genetic alterability, VEEV and CHIKV have great potential for development into biological weapons. Therefore, there is an urgent need to determine possible methods of treatment or prevention. One possibility of prevention lies in determining the structure and biological function of the E3 protein that plays a key role in the infectivity of the viruses. In this research, heavy nitrogen (N15) labeling and nickel-affinity purification were used to purify E3 from VEEV. The labeled E3 was used in nuclear magnetic resonance (NMR) to determine secondary structure, but protein aggregation resulted in poor signal. Although various purification techniques, including cobalt-affinity purification, were used to obtain small amounts of E3 protein from CHIKV, further work is needed in order to obtain enough protein to be used for NMR analysis.

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