Substrate binding activates the designed triple mutant of the colicin E7 metallonuclease
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F14%3A00436908" target="_blank" >RIV/61388963:_____/14:00436908 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1007/s00775-014-1186-6" target="_blank" >http://dx.doi.org/10.1007/s00775-014-1186-6</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1007/s00775-014-1186-6" target="_blank" >10.1007/s00775-014-1186-6</a>
Alternative languages
Result language
angličtina
Original language name
Substrate binding activates the designed triple mutant of the colicin E7 metallonuclease
Original language description
The nuclease domain of colicin E7 (NColE7) cleaves DNA nonspecifically. The active center is a Zn2+-containing HNH motif at the C-terminus. The N-terminal loop is essential for the catalytic activity providing opportunity for allosteric modulation of theenzyme. To identify the key residues responsible for the structural integrity of NColE7, a virtual alanine scan was performed on a semiempirical quantum chemical level within the 25 residue long N-terminal sequence (446-470). Based on the calculations the T454A/K458A/W464A-NColE7 triple mutant (TKW) was expressed and purified. According to the agarose gel electrophoresis experiments and linear dichroism spectra the catalytic activity of the TKW mutant decreased in comparison with wild-type NColE7. Thedistorted structure and weakened Zn2+ binding may account for this as revealed by circular dichroism spectra, mass spectrometry, fluorescence-based thermal analysis and isothermal microcalorimetric titrations. Remarkably, the substrate in
Czech name
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Czech description
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Classification
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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Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2014
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Biological Inorganic Chemistry
ISSN
0949-8257
e-ISSN
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Volume of the periodical
19
Issue of the periodical within the volume
8
Country of publishing house
DE - GERMANY
Number of pages
9
Pages from-to
1295-1303
UT code for WoS article
000345403900005
EID of the result in the Scopus database
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