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Protein purification, crystallization, and structure determination of transcription factor YhaJ in complex with DNT metabolites
- Kim, Myeongbin;
- Kang, Ryun;
- Jeon, Tae Jin;
- Ryu, Seong Eon
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1초록
The microbial transcription factor YhaJ responds to 2,4-dinitrotoluene (DNT) derivatives. Here, we describe steps for overexpression and purification of the protein, characterization for the binding of a DNT derivative methylhydroquinone, and crystallization by using a random seeding technique. We then detail procedures for structure determination by employing the crystal-twin resolving processes. This protocol can also be performed using other DNT derivatives. For complete details on the use and execution of this protocol, please refer to Kim et al.
키워드
Biotechnology and bioengineering; Protein expression and purification; X-ray Crystallography; MACROMOLECULAR STRUCTURES; VALIDATION; INTERFACE; PROGRAM
- 제목
- Protein purification, crystallization, and structure determination of transcription factor YhaJ in complex with DNT metabolites
- 저자
- Kim, Myeongbin; Kang, Ryun; Jeon, Tae Jin; Ryu, Seong Eon
- 발행일
- 2024-06
- 유형
- Article
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- STAR PROTOCOLS
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- 5
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- 2
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- 1 ~ 17