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

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About: Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells Goto Sponge NotDistinct Permalink

An Entity of Type : http://linked.opendata.cz/ontology/domain/vavai/Vysledek, within Data Space : linked.opendata.cz associated with source document(s)

Attributes	Values
rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
rdfs:seeAlso	http://www.sciencedirect.com/science/article/pii/S104659281300257X
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 were successfully 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 uncleaved liposomal DHRS7 were confirmed as enzymes, which reduce carbonyl group of the substrates. 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 were successfully 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 uncleaved liposomal DHRS7 were confirmed as enzymes, which reduce carbonyl group of the substrates. (en)
Title	Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells (en)
skos:prefLabel	Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells Purification and reconstitution of human membrane-bound DHRS7 (SDR34C1) from Sf9 cells (en)
skos:notation	RIV/00216208:11160/14:10281983!RIV15-MSM-11160___
http://linked.open...avai/riv/aktivita	I P S
http://linked.open...avai/riv/aktivity	I, P(EE2.3.20.0235), S
http://linked.open...iv/cisloPeriodika	March
http://linked.open...vai/riv/dodaniDat	2015
http://linked.open...aciTvurceVysledku	Štambergová, Hana Skarka, Adam Wsól, Vladimír Škarydová, Lucie
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Univerzita Karlova v Praze / Farmaceutická fakulta v Hradci Králové
http://linked.open...dnocenehoVysledku	40916
http://linked.open...ai/riv/idVysledku	RIV/00216208:11160/14:10281983
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	Carbonyl reductase activity; Reconstitution; Purification; Solubilisation; DHRS7; Short-chain dehydrogenases/reductases (en)
http://linked.open.../riv/klicoveSlovo	Purification Reconstitution DHRS7 Short-chain dehydrogenases/reductases Carbonyl reductase activity Solubilisation
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[241D9EA30778]
http://linked.open...i/riv/nazevZdroje	Protein Expression and Purification
http://linked.open...in/vavai/riv/obor	CE
http://linked.open...ichTvurcuVysledku	4 (xsd:int)
http://linked.open...cetTvurcuVysledku	4 (xsd:int)
http://linked.open...vavai/riv/projekt	Establishment of Research Team Focused on Experimental and Applied Biopharmacy
http://linked.open...UplatneniVysledku	2014
http://linked.open...v/svazekPeriodika	95
http://linked.open...iv/tvurceVysledku	Skarka, Adam Wsól, Vladimír Škarydová, Lucie Štambergová, Hana
http://linked.open...ain/vavai/riv/wos	000332192900007
issn	1046-5928
number of pages	6 (xsd:int)
http://bibframe.org/vocab/doi	10.1016/j.pep.2013.11.013
http://localhost/t...ganizacniJednotka	11160

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