ID OASD_COMTE Reviewed; 530 AA. AC Q59327; DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot. DT 01-NOV-1996, sequence version 1. DT 13-SEP-2023, entry version 91. DE RecName: Full=3-oxo-5-alpha-steroid 4-dehydrogenase; DE EC=1.3.99.5; DE AltName: Full=Delta 4, 5-alpha steroid dehydrogenase; OS Comamonas testosteroni (Pseudomonas testosteroni). OC Bacteria; Pseudomonadota; Betaproteobacteria; Burkholderiales; OC Comamonadaceae; Comamonas. OX NCBI_TaxID=285; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION AS A DEHYDROGENASE, CATALYTIC RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, SUBCELLULAR RP LOCATION, COFACTOR, AND SUBSTRATE SPECIFICITY. RX PubMed=8655514; DOI=10.1128/jb.178.11.3322-3330.1996; RA Florin C., Kohler T., Grandguillot M., Plesiat P.; RT "Comamonas testosteroni 3-ketosteroid-delta 4(5 alpha)-dehydrogenase: gene RT and protein characterization."; RL J. Bacteriol. 178:3322-3330(1996). RN [2] RP SUBCELLULAR LOCATION. RX PubMed=13673006; DOI=10.1016/s0021-9258(18)69859-x; RA Levy H.R., Talalay P.; RT "Bacterial oxidation of steroids. II. Studies on the enzymatic mechanism of RT ring A dehydrogenation."; RL J. Biol. Chem. 234:2014-2021(1959). CC -!- FUNCTION: Involved in the degradation of steroids having an A:B ring CC fusion in a trans configuration. Catalyzes the elimination of hydrogens CC located at positions 4 and 5 and the introduction of double bonds into CC ring A. {ECO:0000269|PubMed:8655514}. CC -!- CATALYTIC ACTIVITY: CC Reaction=A + a 3-oxo-5alpha-steroid = a 3-oxo-Delta(4)-steroid + AH2; CC Xref=Rhea:RHEA:13805, ChEBI:CHEBI:13193, ChEBI:CHEBI:13601, CC ChEBI:CHEBI:17499, ChEBI:CHEBI:47909; EC=1.3.99.5; CC Evidence={ECO:0000269|PubMed:8655514}; CC -!- CATALYTIC ACTIVITY: CC Reaction=5alpha-androstan-3,17-dione + A = AH2 + androst-4-ene-3,17- CC dione; Xref=Rhea:RHEA:51048, ChEBI:CHEBI:13193, ChEBI:CHEBI:15994, CC ChEBI:CHEBI:16422, ChEBI:CHEBI:17499; EC=1.3.99.5; CC Evidence={ECO:0000269|PubMed:8655514}; CC -!- CATALYTIC ACTIVITY: CC Reaction=5alpha-androst-1-ene-3,17-dione + A = AH2 + androsta-1,4- CC diene-3,17-dione; Xref=Rhea:RHEA:51060, ChEBI:CHEBI:13193, CC ChEBI:CHEBI:17499, ChEBI:CHEBI:40799, ChEBI:CHEBI:133930; CC EC=1.3.99.5; Evidence={ECO:0000269|PubMed:8655514}; CC -!- COFACTOR: CC Name=FAD; Xref=ChEBI:CHEBI:57692; CC Evidence={ECO:0000305|PubMed:8655514}; CC -!- ACTIVITY REGULATION: Inhibition occurs with substrate concentrations CC above 25 uM. {ECO:0000269|PubMed:8655514}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=6.6 uM for 5-alpha-1-androstene-3,17-dione (at 30 degrees Celsius CC and at pH 8.2) {ECO:0000269|PubMed:8655514}; CC KM=16.1 uM for 5-alpha-androstane-3,17-dione (at 30 degrees Celsius CC and at pH 8.2) {ECO:0000269|PubMed:8655514}; CC Vmax=456 nmol/min/mg enzyme with 5-alpha-androstane-3,17-dione as CC substrate (at 30 degrees Celsius and at pH 8.2) CC {ECO:0000269|PubMed:8655514}; CC Vmax=700 nmol/min/mg enzyme with 5-alpha-1-androstene-3,17-dione as CC substrate (at 30 degrees Celsius and at pH 8.2) CC {ECO:0000269|PubMed:8655514}; CC pH dependence: CC Optimum pH is 8. {ECO:0000269|PubMed:8655514}; CC Temperature dependence: CC Optimum temperature is 40 degrees Celsius. CC {ECO:0000269|PubMed:8655514}; CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000305}; Single-pass membrane CC protein {ECO:0000305}. CC -!- SIMILARITY: Belongs to the FAD-dependent oxidoreductase 2 family. CC {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; L23428; AAB08517.1; -; Genomic_DNA. DR PIR; JC6030; JC6030. DR AlphaFoldDB; Q59327; -. DR SMR; Q59327; -. DR KEGG; ag:AAB08517; -. DR BioCyc; MetaCyc:MONOMER-16925; -. DR GO; GO:0016020; C:membrane; IEA:UniProtKB-SubCell. DR GO; GO:0003865; F:3-oxo-5-alpha-steroid 4-dehydrogenase activity; IEA:UniProtKB-EC. DR GO; GO:0047571; F:3-oxosteroid 1-dehydrogenase activity; IDA:UniProtKB. DR GO; GO:0006706; P:steroid catabolic process; IDA:UniProtKB. DR Gene3D; 3.50.50.60; FAD/NAD(P)-binding domain; 1. DR Gene3D; 3.90.700.10; Succinate dehydrogenase/fumarate reductase flavoprotein, catalytic domain; 1. DR InterPro; IPR003953; FAD-binding_2. DR InterPro; IPR036188; FAD/NAD-bd_sf. DR InterPro; IPR027477; Succ_DH/fumarate_Rdtase_cat_sf. DR PANTHER; PTHR43400:SF7; FAD_BINDING_2 DOMAIN-CONTAINING PROTEIN; 1. DR PANTHER; PTHR43400; FUMARATE REDUCTASE; 1. DR Pfam; PF00890; FAD_binding_2; 1. DR PRINTS; PR00411; PNDRDTASEI. DR SUPFAM; SSF51905; FAD/NAD(P)-binding domain; 1. DR SUPFAM; SSF56425; Succinate dehydrogenase/fumarate reductase flavoprotein, catalytic domain; 1. PE 1: Evidence at protein level; KW FAD; Flavoprotein; Lipid metabolism; Membrane; Oxidoreductase; KW Steroid metabolism; Transmembrane; Transmembrane helix. FT CHAIN 1..530 FT /note="3-oxo-5-alpha-steroid 4-dehydrogenase" FT /id="PRO_0000418533" FT TRANSMEM 395..415 FT /note="Helical" FT /evidence="ECO:0000255" FT BINDING 33..62 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000250" SQ SEQUENCE 530 AA; 57289 MW; 5E5D6CEAC0BF2EC1 CRC64; MSNVKKHVST INPVGEVLDV GSADEVQWSD ASDVVVVGWG GAGASAAIEA REQGAEVLVI ERFSGGGASV LSGGVVYAGA VPATRRKPAS RFTEAMTAYL KHEVNGVVSD ETLARFSRDS VTNLNWLEKQ GATFASTMPG YKTSYPADGM YLYYSGNEVV PAYGNPQLLK KPPPRGHRVV AKGQSGAMFF AALQKSTLAH GARTLTQARV QRLVREKDSG RVLGVEVMVL PEGDPRTERH KKLDELVAKS ACIRRRVPRR VAVNVRRSRA RSARSATSVP AKVWCCPLAA ISSIRNCWSM RRYKPGWLTG AAGCDGSGLR LGQSVGGIAQ DLNNISAWRF ITPPSVWPKG LVVNIQGERF CNEQVYGAKL GYEMMEKQGG QAWLIIDSNV RRQAAWQCLF GGLWAFQSMP ALALMYKVAI KGKSVDDLAK KLRMDAAVLQ LQFDRANAPA RGEIEDPLGK SQDMRHEFKG GSLFAIDISI SQKMFPLAVL SLGGLKVNED NGAVIDGAGY DIPGLYAAGV PPLVWLPRVT //