High-Yield Expression in Escherichia coli and Purification of Mouse Ubiquitin-Activating Enzyme E1

• Type: Article in International Scientific Journal
• Date: 2012

Authors

carvalho, af (Author) Other; pinto, mp (Author) Other; grou, cp (Author) Other; vitorino, r (Author) Other; domingues, p (Author) Other; yamao, f (Author) Other; sá-miranda, c (Author) Other; azevedo, je (Author) FCUP

Journal

Title: Molecular Biotechnology

Vol. 51

Pages: 1-8

ISSN: 1073-6085

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 3 Citations

Scopus - 3 Citations

Other information

Authenticus ID: P-002-8RY

DOI: 10.1007/s12033-011-9463-x

Abstract (EN): Research in the ubiquitin field requires large amounts of ubiquitin-activating enzyme (E1) for in vitro ubiquitination assays. Typically, the mammalian enzyme is either isolated from natural sources or produced recombinantly using baculovirus/insect cell protein expression systems. Escherichia coli is seldom used to produce mammalian E1 probably due to the instability and insolubility of this high-molecular mass protein. In this report, we show that 5-10 mg of histidine-tagged mouse E1 can be easily obtained from a 1 l E. coli culture. A low temperature during the protein induction step was found to be critical to obtain an active enzyme.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8