2ISY

Crystal structure of the nickel-activated two-domain iron-dependent regulator (IdeR)

2ISY の概要

• エントリーDOI: 10.2210/pdb2isy/pdb
• 関連するPDBエントリー: 1FX7 1U8R 2ISZ 2IT0
• 分子名称: Iron-dependent repressor ideR, NICKEL (II) ION, PHOSPHATE ION, ... (4 entities in total)
• 機能のキーワード: dna-binding protein, transcription
• 由来する生物種: Mycobacterium tuberculosis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 36167.24

構造登録者

Wisedchaisri, G.,Chou, C.J.,Wu, M.,Roach, C.,Rice, A.E.,Holmes, R.K.,Beeson, C.,Hol, W.G. (登録日: 2006-10-18, 公開日: 2007-02-13, 最終更新日: 2024-11-06)

主引用文献

Wisedchaisri, G.,Chou, C.J.,Wu, M.,Roach, C.,Rice, A.E.,Holmes, R.K.,Beeson, C.,Hol, W.G.
Crystal structures, metal activation, and DNA-binding properties of two-domain IdeR from Mycobacterium tuberculosis
Biochemistry, 46:436-447, 2007

PubMed Abstract: The iron-dependent regulator IdeR is a key transcriptional regulator of iron uptake in Mycobacterium tuberculosis. In order to increase our insight into the role of the SH3-like third domain of this essential regulator, the metal-binding and DNA-binding properties of two-domain IdeR (2D-IdeR) whose SH3-like domain has been truncated were characterized. The equilibrium dissociation constants for Co2+ and Ni2+ activation of 2D-IdeR for binding to the fxbA operator and the DNA-binding affinities of 2D-IdeR in the presence of excess metal ions were estimated using fluorescence spectroscopy. 2D-IdeR binds to fxbA operator DNA with similar affinity as full-length IdeR in the presence of excess metal ion. However, the Ni2+ concentrations required to activate 2D-IdeR for DNA binding appear to be smaller than that for full-length IdeR while the concentration of Co2+ required for activation remains the same. We have determined the crystal structures of Ni2+-activated 2D-IdeR at 1.96 A resolution and its double dimer complex with the mbtA-mbtB operator DNA in two crystal forms at 2.4 A and 2.6 A, the highest resolutions for DNA complexes for any structures of iron-dependent regulator family members so far. The 2D-IdeR-DNA complex structures confirm the specificity of Ser37 and Pro39 for thymine bases and suggest preferential contacts of Gln43 to cytosine bases of the DNA. In addition, our 2D-IdeR structures reveal a remarkable property of the TEV cleavage sequence remaining after removal of the C-terminal His6. This C-terminal tail promotes crystal contacts by forming a beta-sheet with the corresponding tail of neighboring subunits in two unrelated structures of 2D-IdeR, one with and one without DNA. The contact-promoting properties of this C-terminal TEV cleavage sequence may be beneficial for crystallizing other proteins.

PubMed: 17209554
DOI: 10.1021/bi0609826

実験手法

X-RAY DIFFRACTION (1.955 Å)