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

Kód výsledku v 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>

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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

Název v anglickém jazyce

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

Popis výsledku anglicky

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

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

—

Projekt

<a href="/cs/project/LO1302" target="_blank" >LO1302: InterBioMed</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2015

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Inorganic Biochemistry

ISSN

0162-0134

e-ISSN

—

Svazek periodika

151

Číslo periodika v rámci svazku

Oct

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

7

Strana od-do

143-149

Kód UT WoS článku

000367421100017

EID výsledku v databázi Scopus

2-s2.0-84949624093