A new insight into the zinc-dependent DNA-cleavage by the colicin E7 nuclease: a crystallographic and computational study
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A new insight into the zinc-dependent DNA-cleavage by the colicin E7 nuclease : a crystallographic and computational study. / Czene, Anikó; Tóth, Eszter; Németh, Eszter; Otten, Harm; Poulsen, Jens-Christian Navarro; Christensen, Hans Erik Mølager; Rulíšek, Lubomír; Nagata, Kyosuke; Larsen, Sine; Gyurcsik, Béla.
I: Metallomics, Bind 6, Nr. 11, 2014, s. 2090-2099.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - A new insight into the zinc-dependent DNA-cleavage by the colicin E7 nuclease
T2 - a crystallographic and computational study
AU - Czene, Anikó
AU - Tóth, Eszter
AU - Németh, Eszter
AU - Otten, Harm
AU - Poulsen, Jens-Christian Navarro
AU - Christensen, Hans Erik Mølager
AU - Rulíšek, Lubomír
AU - Nagata, Kyosuke
AU - Larsen, Sine
AU - Gyurcsik, Béla
PY - 2014
Y1 - 2014
N2 - The nuclease domain of colicin E7 metallonuclease (NColE7) contains its active centre at the C-terminus. The mutant ΔN4-NColE7-C∗-where the four N-terminal residues including the positively charged K446, R447 and K449 are replaced with eight residues from the GST tag-is catalytically inactive. The crystal structure of this mutant demonstrates that its overall fold is very similar to that of the native NColE7 structure. This implicates the stabilizing effect of the remaining N-terminal sequence on the structure of the C-terminal catalytic site and the essential role of the deleted residues in the mechanism of the catalyzed reaction. Complementary QM/MM calculations on the protein-DNA complexes support the less favourable cleavage by the mutant protein than by NColE7. Furthermore, a water molecule as a possible ligand for the Zn2+-ion is proposed to play a role in the catalytic process. These results suggest that the mechanism of the Zn2+-containing HNH nucleases needs to be further studied and discussed.
AB - The nuclease domain of colicin E7 metallonuclease (NColE7) contains its active centre at the C-terminus. The mutant ΔN4-NColE7-C∗-where the four N-terminal residues including the positively charged K446, R447 and K449 are replaced with eight residues from the GST tag-is catalytically inactive. The crystal structure of this mutant demonstrates that its overall fold is very similar to that of the native NColE7 structure. This implicates the stabilizing effect of the remaining N-terminal sequence on the structure of the C-terminal catalytic site and the essential role of the deleted residues in the mechanism of the catalyzed reaction. Complementary QM/MM calculations on the protein-DNA complexes support the less favourable cleavage by the mutant protein than by NColE7. Furthermore, a water molecule as a possible ligand for the Zn2+-ion is proposed to play a role in the catalytic process. These results suggest that the mechanism of the Zn2+-containing HNH nucleases needs to be further studied and discussed.
U2 - 10.1039/c4mt00195h
DO - 10.1039/c4mt00195h
M3 - Journal article
C2 - 25179124
AN - SCOPUS:84908143969
VL - 6
SP - 2090
EP - 2099
JO - Metallomics
JF - Metallomics
SN - 1756-5901
IS - 11
ER -
ID: 130977630