Fine tuning of the catalytic activity of colicin E7 nuclease domain by systematic N-terminal mutations
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F14%3A00431504" target="_blank" >RIV/61388963:_____/14:00431504 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1002/pro.2497" target="_blank" >http://dx.doi.org/10.1002/pro.2497</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/pro.2497" target="_blank" >10.1002/pro.2497</a>
Alternative languages
Result language
angličtina
Original language name
Fine tuning of the catalytic activity of colicin E7 nuclease domain by systematic N-terminal mutations
Original language description
The nuclease domain of colicin E7 (NColE7) promotes the nonspecific cleavage of nucleic acids at its C-terminal HNH motif. Interestingly, the deletion of four N-terminal residues (446-449 NColE7=KRNK) resulted in complete loss of the enzyme activity. R447A mutation was reported to decrease the nuclease activity, but a detailed analysis of the role of the highly positive and flexible N-terminus is still missing. Here, we present the study of four mutants, with a decreased activity in the following order:NColE7 > KGNK > KGNG similar to GGNK > GGNG. At the same time, the folding, the metal-ion, and the DNA-binding affinity were unaffected by the mutations as revealed by linear and circular dichroism spectroscopy, isothermal calorimetric titrations, and gel mobility shift experiments. Semiempirical quantum chemical calculations and molecular dynamics simulations revealed that K446, K449, and/or the N-terminal amino group are able to approach the active centre in the absence of the other p
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
Protein Science
ISSN
0961-8368
e-ISSN
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Volume of the periodical
23
Issue of the periodical within the volume
8
Country of publishing house
US - UNITED STATES
Number of pages
10
Pages from-to
1113-1122
UT code for WoS article
000339664800010
EID of the result in the Scopus database
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