title: Molecular and biochemical characterisation of human short-chain dehydrogenase/reductase member 3 (DHRS3).
abstract: Dehydrogenase/reductase (SDR family) member 3 (DHRS3), also known as retinal short-chain dehydrogenase/reductase (retSDR1) is a member of SDR16C family. This family is thought to be NADP(H) dependent and to have multiple substrates; however, to date, only all-trans-retinal has been identified as a DHRS3 substrate. The reductive reaction catalysed by DHRS3 seems to be physiological, and recent studies proved the importance of DHRS3 for maintaining suitable retinoic acid levels during embryonic development in vivo. Although it seems that DHRS3 is an important protein, knowledge of the protein and its properties is quite limited, with the majority of information being more than 15 years old. This study aimed to generate a more comprehensive characterisation of the DHRS3 protein. Recombinant enzyme was prepared and demonstrated to be a microsomal, integral-membrane protein with the C-terminus oriented towards the cytosol, consistent with its preference of NADPH as a cofactor. It was determined that DHRS3 also participates in the metabolism of other endogenous compounds, such as androstenedione, estrone, and DL-glyceraldehyde, and in the biotransformation of xenobiotics (e.g., NNK and acetohexamide) in addition to all-trans-retinal. Purified and reconstituted enzyme was prepared for the first time and will be used for further studies. Expression of DHRS3 was shown at the level of both mRNA and protein in the human liver, testis and small intestine. This new information could open other areas of DHRS3 protein research.
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: lundovat@faf.cuni.cz.
authors.forename: Tereza
authors.initials: T
authors.lastname: Lundová
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: lucie.skarydova@faf.cuni.cz.
authors.forename: Lucie
authors.initials: L
authors.lastname: Zemanová
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: malcekob@faf.cuni.cz.
authors.forename: Beata
authors.initials: B
authors.lastname: Malčeková
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: skara3aa@faf.cuni.cz.
authors.forename: Adam
authors.initials: A
authors.lastname: Skarka
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: hana.stambergova@faf.cuni.cz.
authors.forename: Hana
authors.initials: H
authors.lastname: Štambergová
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: havrj4aa@faf.cuni.cz.
authors.forename: Jana
authors.initials: J
authors.lastname: Havránková
authors.affiliation: Institute of Legal Medicine, Faculty of Medicine, Charles University and University Hospital in Hradec Králové, Sokolská 581, 500 05 Hradec Králové, Czech Republic. Electronic address: miroslav.safr@fnhk.cz.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Šafr
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic. Electronic address: vladimir.wsol@faf.cuni.cz.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Wsól
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000818:Animals; D003600:Cytosol / Q000378:metabolism; D064429:Fatty Acid Synthases / Q000378:metabolism*; D006801:Humans; D007421:Intestine, Small / Q000201:enzymology / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera / Q000378:metabolism; D013737:Testis / Q000201:enzymology / Q000378:metabolism; D014212:Tretinoin / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D009249:NADP; D014212:Tretinoin; D000429:Alcohol Oxidoreductases; C036543:short chain trans-2-enoyl-CoA reductase; D009247:NADH, NADPH Oxidoreductases; D064429:Fatty Acid Synthases
keywords: DHRS3; Expression; Membrane topology; Reductase activity; retSDR1