High-Yield Expression of Mouse Aos1-Uba2-Fusion SUMO-Activating Enzyme, mAU, in a Baculovirus-Insect Cell System

Abstract

Small ubiquitin-related modifier (SUMO) research requires large amounts of SUMO-activating enzyme (SUMO-E1) for in vitro SUMOylation assays. In this study the expression and purification of a hexahistidine-tagged mouse Aos1-Uba2-fusion protein (His₆-mAU) in a baculovirus-insect cell system revealed that approximately 1.0 mg of highly-purified His₆-mAU was obtained from 100 mL of Sf9 cell culture. The recombinant protein had SUMO-E1 activity in multiple in vitro SUMOylation assays.