Enhanced Mitogenic Activity of Recombinant Human Vascular Endothelial Growth Factor VEGF(121) Expressed in E. coli Origami B (DE3) with Molecular Chaperones
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F16%3A00467201" target="_blank" >RIV/67985823:_____/16:00467201 - isvavai.cz</a>
Alternative codes found
RIV/61388971:_____/16:00465129 RIV/61389013:_____/16:00465129 RIV/00216208:11310/16:10328400 RIV/00064165:_____/16:10328400
Result on the web
<a href="http://dx.doi.org/10.1371/journal.pone.0163697" target="_blank" >http://dx.doi.org/10.1371/journal.pone.0163697</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1371/journal.pone.0163697" target="_blank" >10.1371/journal.pone.0163697</a>
Alternative languages
Result language
angličtina
Original language name
Enhanced Mitogenic Activity of Recombinant Human Vascular Endothelial Growth Factor VEGF(121) Expressed in E. coli Origami B (DE3) with Molecular Chaperones
Original language description
We describe the production of a highly-active mutant VEGF variant, alpha(2)-PI1-8-VEGF(121), which contains a substrate sequence for factor XIIIa at the aminoterminus designed for incorporation into a fibrin gel. The alpha(2)-PI1-8-VEGF(121) gene was synthesized, cloned into a pET-32a(+) vector and expressed in Escherichia coli Origami B (DE3) host cells. To increase the protein folding and the solubility, the resulting thioredoxin-alpha(2)-PI1-8-VEGF(121) fusion protein was co-expressed with recombinant molecular chaperones GroES/EL encoded by independent plasmid pGro7.The fusion protein was purified from the soluble fraction of cytoplasmic proteins using affinity chromatography. After cleavage of the thioredoxin fusion part with thrombin, the target protein was purified by a second round of affinity chromatography. The yield of purified alpha(2)-PI1-8-VEGF(121) was 1.4 mg per liter of the cell culture. The alpha(2)-PI1-8-VEGF(121) expressed in this work increased the proliferation of endothelial cells 3.9-8.7 times in comparison with commercially-available recombinant VEGF(121). This very high mitogenic activity may be caused by co-expression of the growth factor with molecular chaperones not previously used in VEGF production. At the same time, alpha(2)-PI1-8-VEGF(121) did not elicit considerable inflammatory activation of human endothelial HUVEC cells and human monocyte-like THP-1 cells.
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
FA - Cardiovascular diseases including cardio-surgery
OECD FORD branch
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Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2016
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
PLoS ONE
ISSN
1932-6203
e-ISSN
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Volume of the periodical
11
Issue of the periodical within the volume
10
Country of publishing house
US - UNITED STATES
Number of pages
22
Pages from-to
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UT code for WoS article
000385698100010
EID of the result in the Scopus database
2-s2.0-84991449375