Preorganization of the catalytic Zn2+-binding site in the HNH nuclease motif-A solution study

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F15%3A00455686" target="_blank" >RIV/61388963:_____/15:00455686 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1016/j.jinorgbio.2015.03.017" target="_blank" >http://dx.doi.org/10.1016/j.jinorgbio.2015.03.017</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.jinorgbio.2015.03.017" target="_blank" >10.1016/j.jinorgbio.2015.03.017</a>

Result language
angličtina
Original language name
Preorganization of the catalytic Zn2+-binding site in the HNH nuclease motif-A solution study
Original language description
The structure of the active site in a metalloenzyme can be a key determinant of its metal ion binding affinity and catalytic activity. In this study, the conformational features of the Zn2+-binding HNH motif were investigated by CD-spectroscopy in combination with isothermal microcalorimetric titrations. Various point mutations, including T454A, K458A and W464A, were introduced into the N-terminal loop of the nuclease domain of colicin E7 (NColE7). We show that the folding of the proteins was severely disturbed by the mutation of the tryptophan residue. This points to the importance of W464, being a part of the hydrophobic core located close to the HNH-motif. ITC demonstrated that the Zn2+-binding of the mutants including the W464 site became weak, andaccording to CD-spectroscopic measurements the addition of the metal ion itself cannot fully recover the functional structure. Titrations with Zn2+-ion in the presence and absence of the Im7 protein proved that the structural changes in
Czech name
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Czech description
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Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
CE - Biochemistry
OECD FORD branch
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Project
<a href="/en/project/LO1302" target="_blank" >LO1302: InterBioMed</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2015
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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