The molten-globule residual structure is critical for reflavination of glucose oxidase
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F17%3A00484183" target="_blank" >RIV/61388971:_____/17:00484183 - isvavai.cz</a>
Alternative codes found
RIV/61989592:15310/17:73586491
Result on the web
<a href="http://dx.doi.org/10.1016/j.bpc.2017.08.009" target="_blank" >http://dx.doi.org/10.1016/j.bpc.2017.08.009</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.bpc.2017.08.009" target="_blank" >10.1016/j.bpc.2017.08.009</a>
Alternative languages
Result language
angličtina
Original language name
The molten-globule residual structure is critical for reflavination of glucose oxidase
Original language description
Glucose oxidase (GOX) is a homodimeric glycoprotein with tightly bound one molecule of FAD cofactor per monomer of the protein. GOX has numerous applications, but the preparation of biotechnologically interesting GOX sensors requires a removal of the native FAD cofactor. This process often leads to unwanted irreversible deflavination and, as a consequence, to the low enzyme recovery. Molecular mechanisms of reversible reflavination are poorly understood, our current knowledge is based only on empiric rules, which is clearly insufficient for further development. To develop conceptual understanding of flavin-binding competent states, we studied the effect of deflavination protocols on conformational properties of GOX. After deflavination, the apoform assembles into soluble oligomers with nearly native-like holoform secondary structure but largely destabilized tertiary structure presumambly due to the packing density defects around the vacant flavin binding site. The reflavination is cooperative but not fully efficient, after the binding the flavin cofactor, the protein directly disassembles into native homodimers while the fraction of oligomers remains irreversibly inactivated. Importantly, the effect of Hofineister salts on the conformational properties of GOX and reflavination efficiency indicates that the native-like residual tertiary structure in the molten-globule states favorably supports the reflavination and minimizes the inactivated oligomers. We interpret our results by combining the ligand-induced changes in quaternary structure with salt-sensitive, non-equilibrated conformational selection model. In summary, our work provides the very first steps toward molecular understanding the complexity of the GOX reflavination mechanism.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10606 - Microbiology
Result continuities
Project
—
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Biophysical Chemistry
ISSN
0301-4622
e-ISSN
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Volume of the periodical
230
Issue of the periodical within the volume
NOV 2017
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
10
Pages from-to
74-83
UT code for WoS article
000413390200010
EID of the result in the Scopus database
2-s2.0-85028733236