Abstract
For HER2 overexpression, the ESX transcription factor must interact with both the HER2 promoter and Sur2, a subunit of human mediator complex, using its Ets domain and transactivation domain, respectively. HER2 overexpression is a marker of poor prognosis in various types of cancers. Thus, interfering with the ESX-Sur2 interaction has been suggested as a novel strategy for the treatment of HER2 positive cancers. To design small molecule inhibitors against the ESX-Sur2 interaction, it is necessary to identify the structure of the interface of ESX-Sur2 binding. Therefore, in this study, we determined the optimal conditions for the overexpression and purification of a new version of Sur2, Sur2391-582, which was able to bind to ESX. To stabilize (His) 6-Sur2391-582, various different buffered conditions over a wide range of pH and ionic strengths were examined. The molecular mass of (His)6-Sur2391-582 was determined using mass spectroscopy and size exclusion chromatography. The purified (His)6-Sur 2391-582 protein displayed the same biological properties as that of the Sur2 full-length protein.
Original language | English |
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Pages (from-to) | 364-375 |
Number of pages | 12 |
Journal | Preparative Biochemistry and Biotechnology |
Volume | 43 |
Issue number | 4 |
DOIs | |
State | Published - 11 Mar 2013 |
Bibliographical note
Funding Information:This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology (2009-0066925).
Keywords
- ESX-Sur2 binding interface
- human epidermal growth factor receptor 2
- metazoan-specific subunit of human mediator complexes
- transcription factor