TY - JOUR
T1 - Expression and purification of a soluble ESX-binding core domain of sur2
AU - Jeon, Kyung Hwa
AU - Jun, Kyu Yeon
AU - Kim, Eun Y.
AU - Kwon, Youngjoo
N1 - 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).
PY - 2013/3/11
Y1 - 2013/3/11
N2 - 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.
AB - 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.
KW - ESX-Sur2 binding interface
KW - human epidermal growth factor receptor 2
KW - metazoan-specific subunit of human mediator complexes
KW - transcription factor
UR - http://www.scopus.com/inward/record.url?scp=84875516718&partnerID=8YFLogxK
U2 - 10.1080/10826068.2012.738273
DO - 10.1080/10826068.2012.738273
M3 - Article
C2 - 23464919
AN - SCOPUS:84875516718
SN - 1082-6068
VL - 43
SP - 364
EP - 375
JO - Preparative Biochemistry and Biotechnology
JF - Preparative Biochemistry and Biotechnology
IS - 4
ER -
