2KVF

Structure of the three-Cys2His2 domain of mouse testis zinc finger protein

2KVF の概要

• エントリーDOI: 10.2210/pdb2kvf/pdb
• 関連するPDBエントリー: 2KVG 2KVH
• 分子名称: Zinc finger and BTB domain-containing protein 32, ZINC ION (2 entities in total)
• 機能のキーワード: protein/dna, metal-binding, zinc-finger, transcription
• 由来する生物種: Mus musculus (mouse)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 3413.33

構造登録者

Chou, C.-C.,Lou, Y.-C.,Chen, C. (登録日: 2010-03-15, 公開日: 2010-04-14, 最終更新日: 2024-05-01)

主引用文献

Chou, C.-C.,Lou, Y.-C.,Tang, T.K.,Chen, C.
Structure and DNA binding characteristics of the three-Cys(2)His(2) domain of mouse testis zinc finger protein.
Proteins, 78:2202-2212, 2010

PubMed Abstract: The C-terminal three-Cys(2)His(2) zinc-finger domain (TZD) of mouse testis zinc-finger protein binds to the 5'-TGTACAGTGT-3' at the Aie1 (aurora-C) promoter with high specificity. Interestingly, the primary sequence of TZD is unique, possessing two distinct linkers, TGEKP and GAAP, and distinct residues at presumed DNA binding sites at each finger, especially finger 3. A K(d) value of approximately 10(-8) M was obtained from surface plasmon resonance analysis for the TZD-DNA complex. NMR structure of the free TZD showed that each zinc finger forms a typical beta beta alpha fold. On binding to DNA, chemical shift perturbations and the R(2) transverse relaxation rate in finger 3 are significantly smaller than those in fingers 1 and 2, which indicates that the DNA binding affinity in finger 3 is weaker. Furthermore, the shift perturbations between TZD in complex with the cognate DNA and its serial mutants revealed that both ADE7 and CYT8, underlined in 5'-ATATGTACAGTGTTAT-3', are critical in specific binding, and the DNA binding in finger 3 is sequence independent. Remarkably, the shift perturbations in finger 3 on the linker mutation of TZD (GAAP mutated to TGEKP) were barely detected, which further indicates that finger 3 does not play a critical role in DNA sequence-specific recognition. The complex model showed that residues important for DNA binding are mainly located on positions -1, 2, 3, and 6 of alpha-helices in fingers 1 and 2. The DNA sequence and nonsequence-specific bindings occurring simultaneously in TZD provide valuable information for better understanding of protein-DNA recognition.

PubMed: 20544958
DOI: 10.1002/prot.22732

実験手法

SOLUTION NMR