学術雑誌論文

学術雑誌論文

text

PLOS ONE. 2014, 9(6), p. e98554

NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP

PLOS

英語

text

2014-06

https://doi.org/10.1371/journal.pone.0098554

Mizushima R, Kim JY, Suetake I, Tanaka H, Takai T, et al. (2014) NMR Characterization of the Interaction of the Endonuclease Domain of MutL withDivalent Metal Ions and ATP. PLoS ONE 9(6): e98554. doi:10.1371/journal.pone.0098554

MutL is a multi-domain protein comprising an N-terminal ATPase domain (NTD) and C-terminal dimerization domain (CTD), connected with flexible linker regions, that plays a key role in DNA mismatch repair. To expand understanding of the regulation mechanism underlying MutL endonuclease activity, our NMR-based study investigated interactions between the CTD of MutL, derived from the hyperthermophilic bacterium Aquifex aeolicus (aqMutL-CTD), and putative binding molecules. Chemical shift perturbation analysis with the model structure of aqMutL-CTD and circular dichroism results revealed that tight Zn2+ binding increased thermal stability without changing secondary structures to function at high temperatures. Peak intensity analysis exploiting the paramagnetic relaxation enhancement effect indicated the binding site for Mn2+, which shared binding sites for Zn2+. The coexistence of these two metal ions appears to be important for the function of MutL. Chemical shift perturbation analysis revealed a novel ATP binding site in aqMutL-CTD. A docking simulation incorporating the chemical shift perturbation data provided a putative scheme for the intermolecular interactions between aqMutL-CTD and ATP. We proposed a simple and understandable mechanical model for the regulation of MutL endonuclease activity in MMR based on the relative concentrations of ATP and CTD through ATP binding-regulated interdomain interactions between CTD and NTD.

© 2014 Mizushima et al. This article is licensed under a Creative Commons Attribution 4.0 International License.

71261

NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP

PLOS

英語

2014-06

PLOS ONE

9

6

e98554

https://doi.org/10.1371/journal.pone.0098554

https://hdl.handle.net/11094/71261