3QN2

Structure-based design of a disulfide-linked oligomeric form of the Simian Virus 40 (SV40) large T antigen DNA binding domain

3QN2 の概要

• エントリーDOI: 10.2210/pdb3qn2/pdb
• 関連するPDBエントリー: 2FUF
• 分子名称: Large T antigen, CITRATE ANION (3 entities in total)
• 機能のキーワード: origin binding domain, dna replication, hydrolase
• 由来する生物種: Simian virus 40 (SV40)
• 細胞内の位置: Host nucleus: P03070
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15515.69

構造登録者

Meinke, G.,Bullock, P.A. (登録日: 2011-02-07, 公開日: 2011-06-01, 最終更新日: 2024-11-27)

主引用文献

Meinke, G.,Phelan, P.,Fradet-Turcotte, A.,Archambault, J.,Bullock, P.A.
Structure-based design of a disulfide-linked oligomeric form of the simian virus 40 (SV40) large T antigen DNA-binding domain.
Acta Crystallogr.,Sect.D, 67:560-567, 2011

PubMed Abstract: The modular multifunctional protein large T antigen (T-ag) from simian virus 40 orchestrates many of the events needed for replication of the viral double-stranded DNA genome. This protein assembles into single and double hexamers on specific DNA sequences located at the origin of replication. This complicated process begins when the origin-binding domain of large T antigen (T-ag ODB) binds the GAGGC sequences in the central region (site II) of the viral origin of replication. While many of the functions of purified T-ag OBD can be studied in isolation, it is primarily monomeric in solution and cannot assemble into hexamers. To overcome this limitation, the possibility of engineering intermolecular disulfide bonds in the origin-binding domain which could oligomerize in solution was investigated. A recent crystal structure of the wild-type T-ag OBD showed that this domain forms a left-handed spiral in the crystal with six subunits per turn. Therefore, we analyzed the protein interface of this structure and identified two residues that could potentially support an intermolecular disulfide bond if changed to cysteines. SDS-PAGE analysis established that the mutant T-ag OBD formed higher oligomeric products in a redox-dependent manner. In addition, the 1.7 Å resolution crystal structure of the engineered disulfide-linked T-ag OBD is reported, which establishes that oligomerization took place in the expected manner.

PubMed: 21636896
DOI: 10.1107/S0907444911014302

実験手法

X-RAY DIFFRACTION (1.661 Å)