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Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11160%2F14%3A10281983" target="_blank" >RIV/00216208:11160/14:10281983 - isvavai.cz</a>

  • Result on the web

    <a href="http://www.sciencedirect.com/science/article/pii/S104659281300257X" target="_blank" >http://www.sciencedirect.com/science/article/pii/S104659281300257X</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.1016/j.pep.2013.11.013" target="_blank" >10.1016/j.pep.2013.11.013</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells

  • Original language description

    Dehydrogenase/reductase SDR family member 7 (DHRS7, SDR34C1, retSDR4) is one of the many endoplasmic reticulum bound members of the SDR superfamily. Preliminary results indicate its potential significance in human metabolism. DHRS7 containing TEV-cleavable His(10) and FLAG-tag expressed in the Sf9 cell line was solubilised, purified, and reconstituted into liposomes to enable the improved characterisation of this enzyme in the future. Igepal CA-630 was determined to be the best detergent for the solubilisation process. The solubilised DHRS7 was purified using affinity chromatography, and the purified enzyme was subjected to TEV cleavage of the affinity tags and then repurified using subtractive Ni-IMAC. The cleaved and uncleaved versions of DHRS7 weresuccessfully reconstituted into liposomes. In addition, using tobacco specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) as the substrate, the cleaved liposomal DHRS7 was found to be inactive, whereas the pure and un

  • Czech name

  • Czech description

Classification

  • Type

    J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

  • CEP classification

    CE - Biochemistry

  • OECD FORD branch

Result continuities

  • Project

    <a href="/en/project/EE2.3.20.0235" target="_blank" >EE2.3.20.0235: Establishment of Research Team Focused on Experimental and Applied Biopharmacy</a><br>

  • Continuities

    P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach<br>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 Expression and Purification

  • ISSN

    1046-5928

  • e-ISSN

  • Volume of the periodical

    95

  • Issue of the periodical within the volume

    March

  • Country of publishing house

    US - UNITED STATES

  • Number of pages

    6

  • Pages from-to

    44-49

  • UT code for WoS article

    000332192900007

  • EID of the result in the Scopus database