N-glycosylation of tomato nuclease TBN1 produced in N. benthamiana and its effect on the enzyme activity
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F18%3A00497108" target="_blank" >RIV/86652036:_____/18:00497108 - isvavai.cz</a>
Alternative codes found
RIV/60077344:_____/18:00497108 RIV/60461373:22330/18:43915976
Result on the web
<a href="http://dx.doi.org/10.1016/j.plantsci.2018.08.011" target="_blank" >http://dx.doi.org/10.1016/j.plantsci.2018.08.011</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.plantsci.2018.08.011" target="_blank" >10.1016/j.plantsci.2018.08.011</a>
Alternative languages
Result language
angličtina
Original language name
N-glycosylation of tomato nuclease TBN1 produced in N. benthamiana and its effect on the enzyme activity
Original language description
A unique analysis of an enzyme activity versus structure modification of the tomato nuclease R-TBN1 is presented. R-TBN1, the non-specific nuclease belonging to the S1-P1 nuclease family, was recombinantly produced in N. benthamiana. The native structure is posttranslationally modified by N-glycosylation at three sites. In this work, it was found that this nuclease is modified by high-mannose type N-glycosylation with a certain degree of macro-and microheterogeneity. To monitor the role of N-glycosylation in its activity, hypo-and hyperglycosylated nuclease mutants, R-TBN1 digested by alpha-mannosidase, and R-TBN1 deglycosylated by PNGase F were prepared. Deglycosylated R-TBN1 and mutant N94D/N112D were virtually inactive. Compared to R-TBN1 wt, both N94D and N112D mutants showed about 60% and 10% of the activity, respectively, while the N186D, D36S, and D36S/E104 N mutants were equally or even more active than R-TBN1 wt. The partial demannosylation of R-TBN1 did not affect the nuclease aktivity, moreover, a little shift in substrate specificity was observed. The results show two facts: 1) which sites must be occupied by a glycan for the proper folding and stability and 2) how N. benthamiana glycosylates the foreign nuclease. At the same time, the modifications can be interesting in designing the nuclease activity or specificity through its glycosylation.
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
10608 - Biochemistry and molecular biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2018
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Plant Science
ISSN
0168-9452
e-ISSN
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Volume of the periodical
276
Issue of the periodical within the volume
NOV 2018
Country of publishing house
IE - IRELAND
Number of pages
10
Pages from-to
152-161
UT code for WoS article
000449310500015
EID of the result in the Scopus database
2-s2.0-85052477021