Kanemaru Ayumi, Saitoh Hisato
Faculty of Sciences, Kumamoto University, Kumamoto 860-8555, Japan.
Biosci Biotechnol Biochem. 2013;77(7):1575-8. doi: 10.1271/bbb.130070. Epub 2013 Jul 7.
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.
小泛素相关修饰物（SUMO）研究需要大量的SUMO激活酶（SUMO-E1）用于体外SUMO化分析。在本研究中，杆状病毒-昆虫细胞系统中六聚组氨酸标签化的小鼠Aos1-Uba2融合蛋白（His6-mAU）的表达和纯化表明，从100 mL Sf9细胞培养物中可获得约1.0 mg的高度纯化的His6-mAU。该重组蛋白在多种体外SUMO化分析中具有SUMO-E1活性。
