KSHB_RHORH
ID KSHB_RHORH Reviewed; 351 AA.
AC B6V6V6;
DT 30-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 16-DEC-2008, sequence version 1.
DT 03-AUG-2022, entry version 53.
DE RecName: Full=3-ketosteroid-9-alpha-monooxygenase, ferredoxin reductase component {ECO:0000303|PubMed:19561185};
DE AltName: Full=3-ketosteroid-9-alpha-hydroxylase, ferredoxin reductase component {ECO:0000303|PubMed:19561185};
DE Short=KSH {ECO:0000303|PubMed:19561185};
DE AltName: Full=Androsta-1,4-diene-3,17-dione 9-alpha-hydroxylase {ECO:0000303|PubMed:19561185};
DE EC=1.14.15.30 {ECO:0000269|PubMed:19561185};
DE AltName: Full=Rieske-type oxygenase {ECO:0000305|PubMed:19561185};
DE Short=RO {ECO:0000305|PubMed:19561185};
GN Name=kshB {ECO:0000303|PubMed:19561185};
OS Rhodococcus rhodochrous.
OC Bacteria; Actinobacteria; Corynebacteriales; Nocardiaceae; Rhodococcus.
OX NCBI_TaxID=1829;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, ACTIVITY REGULATION, SUBUNIT, AND
RP SUBSTRATE SPECIFICITY.
RC STRAIN=DSM 43269;
RX PubMed=19561185; DOI=10.1128/aem.00066-09;
RA Petrusma M., Dijkhuizen L., van der Geize R.;
RT "Rhodococcus rhodochrous DSM 43269 3-ketosteroid 9alpha-hydroxylase, a two-
RT component iron-sulfur-containing monooxygenase with subtle steroid
RT substrate specificity.";
RL Appl. Environ. Microbiol. 75:5300-5307(2009).
CC -!- FUNCTION: Probably involved in the degradation of cholesterol. In
CC vitro, catalyzes the introduction of a 9alpha-hydroxyl moiety into the
CC ring B of 3-ketosteroid substrates such as 1,4-androstadiene-3,17-dione
CC (ADD), 4-androstene-3,17-dione (AD), 4-androstene-17beta-ol-3-one
CC (testosterone), 4-pregnene-3,20-dione (progesterone), 19-nor-4-
CC androstene-3,17-dione (nordion), 1-(5alpha)-androstene-3,17-dione,
CC 5alpha-androstane-3,17-dione and 5beta-androstane-3,17-dione. KSH has
CC the highest activity with 3-keto-Delta4 steroid substrates.
CC {ECO:0000269|PubMed:19561185}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=androsta-1,4-diene-3,17-dione + 2 H(+) + O2 + 2 reduced [2Fe-
CC 2S]-[ferredoxin] = 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + H2O
CC + 2 oxidized [2Fe-2S]-[ferredoxin]; Xref=Rhea:RHEA:32199, Rhea:RHEA-
CC COMP:10000, Rhea:RHEA-COMP:10001, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738, ChEBI:CHEBI:40799, ChEBI:CHEBI:63641;
CC EC=1.14.15.30; Evidence={ECO:0000269|PubMed:19561185};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:19561185};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:19561185};
CC -!- COFACTOR:
CC Name=[2Fe-2S] cluster; Xref=ChEBI:CHEBI:190135;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU00465};
CC Note=Binds 1 2Fe-2S cluster. {ECO:0000255|PROSITE-ProRule:PRU00465};
CC -!- ACTIVITY REGULATION: KSH activity is completely inhibited by zinc ions.
CC KshB is specifically inhibited by Cu(2+) ions.
CC {ECO:0000269|PubMed:19561185}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=10 uM for 19-nor-AD {ECO:0000269|PubMed:19561185};
CC KM=23 uM for 5alpha-androstane-3,17-dione
CC {ECO:0000269|PubMed:19561185};
CC KM=33 uM for 5beta-androstane-3,17-dione
CC {ECO:0000269|PubMed:19561185};
CC pH dependence:
CC Optimum pH is 7. {ECO:0000269|PubMed:19561185};
CC Temperature dependence:
CC Optimum temperature is 33 degrees Celsius.
CC {ECO:0000269|PubMed:19561185};
CC -!- PATHWAY: Steroid metabolism; cholesterol degradation.
CC {ECO:0000305|PubMed:19561185}.
CC -!- SUBUNIT: The two-component system 3-ketosteroid-9-alpha-monooxygenase
CC is composed of an oxygenase component KshA and a reductase component
CC KshB. {ECO:0000269|PubMed:19561185}.
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DR EMBL; FJ238096; ACI62781.1; -; Genomic_DNA.
DR AlphaFoldDB; B6V6V6; -.
DR SMR; B6V6V6; -.
DR BioCyc; MetaCyc:MON-16922; -.
DR UniPathway; UPA01058; -.
DR GO; GO:0051537; F:2 iron, 2 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0036200; F:3-ketosteroid 9-alpha-monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0071949; F:FAD binding; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0006707; P:cholesterol catabolic process; NAS:UniProtKB.
DR CDD; cd00207; fer2; 1.
DR Gene3D; 3.10.20.30; -; 1.
DR Gene3D; 3.40.50.80; -; 1.
DR InterPro; IPR036010; 2Fe-2S_ferredoxin-like_sf.
DR InterPro; IPR001041; 2Fe-2S_ferredoxin-type.
DR InterPro; IPR006058; 2Fe2S_fd_BS.
DR InterPro; IPR012675; Beta-grasp_dom_sf.
DR InterPro; IPR008333; Cbr1-like_FAD-bd_dom.
DR InterPro; IPR017927; FAD-bd_FR_type.
DR InterPro; IPR001709; Flavoprot_Pyr_Nucl_cyt_Rdtase.
DR InterPro; IPR039261; FNR_nucleotide-bd.
DR InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR Pfam; PF00970; FAD_binding_6; 1.
DR Pfam; PF00111; Fer2; 1.
DR Pfam; PF00175; NAD_binding_1; 1.
DR PRINTS; PR00371; FPNCR.
DR SUPFAM; SSF52343; SSF52343; 1.
DR SUPFAM; SSF54292; SSF54292; 1.
DR SUPFAM; SSF63380; SSF63380; 1.
DR PROSITE; PS00197; 2FE2S_FER_1; 1.
DR PROSITE; PS51085; 2FE2S_FER_2; 1.
DR PROSITE; PS51384; FAD_FR; 1.
PE 1: Evidence at protein level;
KW 2Fe-2S; Cholesterol metabolism; FAD; Flavoprotein; Iron; Iron-sulfur;
KW Lipid degradation; Lipid metabolism; Metal-binding; Oxidoreductase;
KW Steroid metabolism; Sterol metabolism.
FT CHAIN 1..351
FT /note="3-ketosteroid-9-alpha-monooxygenase, ferredoxin
FT reductase component"
FT /id="PRO_0000438404"
FT DOMAIN 10..116
FT /note="FAD-binding FR-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT DOMAIN 264..351
FT /note="2Fe-2S ferredoxin-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT BINDING 300
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT BINDING 305
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT BINDING 308
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT BINDING 338
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
SQ SEQUENCE 351 AA; 37887 MW; F146084969AD4B2E CRC64;
MTTVEVPHGS RSVILTVSAV VEETADTRSI VFAVPDELRD KFAYRPGQFL TLRIPSDRTG
SVARCYSLAS SPFTDDAPKV TVKRTSDGYG SNWLCDNIAT GQTLEVLPPA GVFTPKSLDH
DFLLFGAGSG ITPVISILKS ALTQGGGKVV LVYANRDEKS VIFAEELRAL AEKYPTRLTV
VHWLESVQGL PTADQLAAIA APYESYEAFM CGPGPFMDTV HQALNTVGMP RARVHAEVFN
SLSGDPFADQ APVEVSDEDA ADAATVEVEL DGEVHKLSWP RKQTLVDIML AKGIDVPYSC
QEGECGSCAC TVLEGKVEME NCDVLDPEDI EAGYILGCQA RPVTDHLKIE F
