title: Expression and purification of a soluble ESX-binding core domain of SUR2.
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-Sur2391-582 protein displayed the same biological properties as that of the Sur2 full-length protein.
authors: College of Pharmacy, Ewha Womans University, Seoul, Korea.
authors: Kyung-Hwa
authors: KH
authors: Jeon
authors: Kyu-Yeon
authors: KY
authors: Jun
authors: Eun Young
authors: EY
authors: Kim
authors: Youngjoo
authors: Y
authors: Kwon
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D056892:Mediator Complex / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D050783:Proto-Oncogene Proteins c-ets; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection
chemical_list: D004268:DNA-Binding Proteins; C105529:ELF3 protein, human; C492159:MED23 protein, human; D056892:Mediator Complex; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors