KSHA4_RHORH
ID KSHA4_RHORH Reviewed; 378 AA.
AC B6V6V5;
DT 30-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 16-DEC-2008, sequence version 1.
DT 03-AUG-2022, entry version 45.
DE RecName: Full=3-ketosteroid-9-alpha-monooxygenase, oxygenase component {ECO:0000303|PubMed:19561185};
DE AltName: Full=3-ketosteroid-9-alpha-hydroxylase, oxygenase 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, ECO:0000305|PubMed:21642460};
DE AltName: Full=Rieske-type oxygenase {ECO:0000305|PubMed:19561185};
DE Short=RO {ECO:0000305|PubMed:19561185};
GN Name=kshA {ECO:0000303|PubMed:21642460};
GN Synonyms=kshA4 {ECO:0000303|PubMed:21642460};
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).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, INDUCTION,
RP AND SUBSTRATE SPECIFICITY.
RC STRAIN=DSM 43269;
RX PubMed=21642460; DOI=10.1128/jb.00274-11;
RA Petrusma M., Hessels G., Dijkhuizen L., van der Geize R.;
RT "Multiplicity of 3-ketosteroid-9alpha-hydroxylase enzymes in Rhodococcus
RT rhodochrous DSM43269 for specific degradation of different classes of
RT steroids.";
RL J. Bacteriol. 193:3931-3940(2011).
CC -!- FUNCTION: In vitro, catalyzes the introduction of a 9alpha-hydroxyl
CC moiety into the ring B of 3-ketosteroid substrates such as 1,4-
CC androstadiene-3,17-dione (ADD), 4-androstene-3,17-dione (AD), 4-
CC androstene-17beta-ol-3-one (testosterone), 4-pregnene-3,20-dione
CC (progesterone), 19-nor-4-androstene-3,17-dione (nordion), 1-(5alpha)-
CC androstene-3,17-dione, 5alpha-androstane-3,17-dione and 5beta-
CC androstane-3,17-dione (PubMed:19561185, PubMed:21642460). KSH has the
CC highest activity with 3-keto-delta4 steroid substrates
CC (PubMed:19561185). {ECO:0000269|PubMed:19561185,
CC ECO:0000269|PubMed:21642460}.
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 ECO:0000305|PubMed:21642460};
CC -!- COFACTOR:
CC Name=[2Fe-2S] cluster; Xref=ChEBI:CHEBI:190135;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU00628,
CC ECO:0000269|PubMed:19561185};
CC Note=Binds 1 [2Fe-2S] cluster per subunit. {ECO:0000255|PROSITE-
CC ProRule:PRU00628, ECO:0000269|PubMed:19561185};
CC -!- COFACTOR:
CC Name=Fe cation; Xref=ChEBI:CHEBI:24875;
CC Evidence={ECO:0000269|PubMed:19561185};
CC Note=Binds 1 Fe cation. {ECO:0000269|PubMed:19561185};
CC -!- ACTIVITY REGULATION: KSH activity is completely inhibited by zinc ions.
CC KshA is specifically inhibited by Fe(3+), Co(2+), Zn(2+) and Ni(2+)
CC ions. {ECO:0000269|PubMed:19561185}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=10 uM for nordion {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 -!- SUBUNIT: Homotrimer (By similarity). The two-component system 3-
CC ketosteroid-9-alpha-monooxygenase is composed of an oxygenase component
CC KshA and a reductase component KshB (PubMed:19561185).
CC {ECO:0000250|UniProtKB:F1CMY8, ECO:0000269|PubMed:19561185}.
CC -!- INDUCTION: By cholic acid. {ECO:0000269|PubMed:21642460}.
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; FJ238095; ACI62780.1; -; Genomic_DNA.
DR EMBL; HQ425876; ADY18323.1; -; Genomic_DNA.
DR AlphaFoldDB; B6V6V5; -.
DR SMR; B6V6V5; -.
DR BioCyc; MetaCyc:MON-16921; -.
DR BRENDA; 1.14.15.30; 5395.
DR GO; GO:0051537; F:2 iron, 2 sulfur cluster binding; IDA:UniProtKB.
DR GO; GO:0036200; F:3-ketosteroid 9-alpha-monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IDA:UniProtKB.
DR GO; GO:0008203; P:cholesterol metabolic process; IEA:InterPro.
DR GO; GO:0016042; P:lipid catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 2.102.10.10; -; 1.
DR InterPro; IPR045605; KshA-like_C.
DR InterPro; IPR017941; Rieske_2Fe-2S.
DR InterPro; IPR036922; Rieske_2Fe-2S_sf.
DR Pfam; PF19298; KshA_C; 1.
DR Pfam; PF00355; Rieske; 1.
DR SUPFAM; SSF50022; SSF50022; 1.
DR PROSITE; PS51296; RIESKE; 1.
PE 1: Evidence at protein level;
KW 2Fe-2S; Iron; Iron-sulfur; Lipid degradation; Lipid metabolism;
KW Metal-binding; Oxidoreductase; Steroid metabolism.
FT CHAIN 1..378
FT /note="3-ketosteroid-9-alpha-monooxygenase, oxygenase
FT component"
FT /id="PRO_0000438402"
FT DOMAIN 26..128
FT /note="Rieske"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 67
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 69
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 86
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 89
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 175
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000250|UniProtKB:F1CMY8"
FT BINDING 181
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000250|UniProtKB:F1CMY8"
FT BINDING 186
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000250|UniProtKB:F1CMY8"
FT BINDING 305
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000250|UniProtKB:F1CMY8"
SQ SEQUENCE 378 AA; 43507 MW; 5D6201C652F417B4 CRC64;
MTVPQERIEI RNIDPGTNPT RFARGWHCIG LAKDFRDGKP HQVKVFGTDL VVFADTAGKL
HVLDAFCRHM GGNLARGEIK GDTIACPFHD WRWNGQGRCE AVPYARRTPK LGRTKAWTTM
ERNGVLFVWH CPQGSEPTPE LAIPEIEGYE DGQWSDWTWT TIHVEGSHCR EIVDNVVDMA
HFFYVHFQMP EYFKNVFDGH IAGQHMRSYG RDDIKTGVQM DLPEAQTISD AFYYGPSFML
DTIYTVSEGT TIESKLINCH YPVTNNSFVL QFGTIVKKIE GMSEEQAAEM ATMFTDGLEE
QFAQDIEIWK HKSRIENPLL TEEDGPVYQL RRWYNQFYVD LEDVTPDMTQ RFEFEVDTSR
ALESWHKEVE ENLAGTAE
