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

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 (His 6 -mAU) in a baculovirus-insect cell system reveale...

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Published inBioscience, biotechnology, and biochemistry Vol. 77; no. 7; pp. 1575 - 1578
Main Authors KANEMARU, Ayumi, SAITOH, Hisato
Format Journal Article
LanguageEnglish
Published England Japan Society for Bioscience, Biotechnology, and Agrochemistry 2013
Oxford University Press
Subjects
Animals
Assaying
Baculoviridae - genetics
baculovirus-insect cell system
Biochemistry
Biotechnology
Enzymes
Gene Expression
Genetic Engineering - methods
In vitro testing
Mice
post-translational protein modification
protein expression
Proteins
Purification
Recombinant
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Sf9 Cells
small ubiquitin-related modifier (SUMO)
Spodoptera
SUMO-activating enzyme (SUMO-E1)
Sumoylation
Ubiquitin-Activating Enzymes - genetics
Ubiquitin-Activating Enzymes - metabolism
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Summary: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 (His 6 -mAU) in a baculovirus-insect cell system revealed that approximately 1.0 mg of highly-purified His 6 -mAU was obtained from 100 mL of Sf9 cell culture. The recombinant protein had SUMO-E1 activity in multiple in vitro SUMOylation assays.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.130070