Crystal structural analysis and metal-dependent stability and activity studies of the ColE7 endonuclease domain in complex with DNA/Zn2+ or inhibitor/Ni2+.

@article{Doudeva2006CrystalSA,
  title={Crystal structural analysis and metal-dependent stability and activity studies of the ColE7 endonuclease domain in complex with DNA/Zn2+ or inhibitor/Ni2+.},
  author={Lyudmila G. Doudeva and Hsinchin Huang and Kuo-Chiang Hsia and Zhonghao Shi and Chia-Lung Li and Yongliang Shen and Y Cheng and Hanna S. Yuan},
  journal={Protein science : a publication of the Protein Society},
  year={2006},
  volume={15 2},
  pages={269-80}
}
The nuclease domain of ColE7 (N-ColE7) contains an H-N-H motif that folds in a beta beta alpha-metal topology. Here we report the crystal structures of a Zn2+-bound N-ColE7 (H545E mutant) in complex with a 12-bp duplex DNA and a Ni2+-bound N-ColE7 in complex with the inhibitor Im7 at a resolution of 2.5 A and 2.0 A, respectively. Metal-dependent cleavage assays showed that N-ColE7 cleaves double-stranded DNA with a single metal ion cofactor, Ni2+, Mg2+, Mn2+, and Zn2+. ColE7 purified from… CONTINUE READING