Expression and purification of a soluble ESX-binding core domain of sur2

Kyung Hwa Jeon; Kyu Yeon Jun; Eun Y. Kim; Youngjoo Kwon

doi:10.1080/10826068.2012.738273

Expression and purification of a soluble ESX-binding core domain of sur2

Kyung Hwa Jeon, Kyu Yeon Jun, Eun Y. Kim, Youngjoo Kwon

Department of Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

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, Sur2_391-582, which was able to bind to ESX. To stabilize (His) ₆-Sur2_391-582, various different buffered conditions over a wide range of pH and ionic strengths were examined. The molecular mass of (His)₆-Sur_2391-582 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
Pages (from-to)	364-375
Number of pages	12
Journal	Preparative Biochemistry and Biotechnology
Volume	43
Issue number	4
DOIs	https://doi.org/10.1080/10826068.2012.738273
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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1080/10826068.2012.738273

Cite this

@article{e25cfb5d77bf40b4a191e8b2c16163a7,

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-Sur 2391-582 protein displayed the same biological properties as that of the Sur2 full-length protein.",

keywords = "ESX-Sur2 binding interface, human epidermal growth factor receptor 2, metazoan-specific subunit of human mediator complexes, transcription factor",

author = "Jeon, {Kyung Hwa} and Jun, {Kyu Yeon} and Kim, {Eun Y.} and Youngjoo Kwon",

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).",

year = "2013",

month = mar,

day = "11",

doi = "10.1080/10826068.2012.738273",

language = "English",

volume = "43",

pages = "364--375",

journal = "Preparative Biochemistry and Biotechnology",

issn = "1082-6068",

publisher = "Taylor and Francis Ltd.",

number = "4",

}

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 -

Expression and purification of a soluble ESX-binding core domain of sur2

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Fingerprint

Cite this