KSHA5_RHORH
ID KSHA5_RHORH Reviewed; 390 AA.
AC F1CMY8;
DT 30-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 03-MAY-2011, sequence version 1.
DT 03-AUG-2022, entry version 38.
DE RecName: Full=3-ketosteroid-9-alpha-monooxygenase, oxygenase component {ECO:0000303|PubMed:21642460};
DE AltName: Full=3-ketosteroid-9-alpha-hydroxylase, oxygenase component {ECO:0000303|PubMed:21642460};
DE Short=KSH {ECO:0000303|PubMed:21642460};
DE AltName: Full=Androsta-1,4-diene-3,17-dione 9-alpha-hydroxylase {ECO:0000303|PubMed:25049233};
DE EC=1.14.15.30 {ECO:0000269|PubMed:25049233, ECO:0000305|PubMed:21642460};
DE AltName: Full=Rieske-type oxygenase {ECO:0000305|PubMed:25049233};
DE Short=RO {ECO:0000305|PubMed:25049233};
GN Name=kshA {ECO:0000303|PubMed:21642460};
GN Synonyms=kshA5 {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, 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).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH IRON ION AND
RP IRON-SULFUR (2FE-2S), FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, COFACTOR, PATHWAY, SUBUNIT, AND SUBSTRATE SPECIFICITY.
RX PubMed=25049233; DOI=10.1074/jbc.m114.575886;
RA Penfield J.S., Worrall L.J., Strynadka N.C., Eltis L.D.;
RT "Substrate specificities and conformational flexibility of 3-ketosteroid
RT 9alpha-hydroxylases.";
RL J. Biol. Chem. 289:25523-25536(2014).
CC -!- FUNCTION: Probably involved in the degradation of cholesterol
CC (PubMed:21642460). In vitro, catalyzes the introduction of a 9alpha-
CC hydroxyl moiety into the ring B of 3-ketosteroid substrates such as
CC 1,4-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, 1-(5alpha)-androstene-
CC 3,17-dione, 5alpha-androstane-3,17-dione, 5beta-androstane-3,17-dione,
CC 5alpha-androstane-17beta-ol-3-one (stanolon), 11beta-hydrocortisone, 3-
CC oxo-23,24-bisnorcholesta-4-en-22-oate (4-BNC), 23,24-bisnorcholesta-4-
CC ene-22-oate, 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate (1,4-BNC) and
CC 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-coenzyme A thioester (1,4-
CC BNC-CoA) (PubMed:21642460, PubMed:25049233). KshA5 has the broadest
CC substrate range without a clear substrate preference and is active with
CC Delta-4, Delta-1,4, 5alpha-H and 5beta-H steroids, as well as with
CC steroids having bulky aliphatic side chains and an isopropionyl side
CC chain at C17. {ECO:0000269|PubMed:21642460,
CC ECO:0000269|PubMed:25049233}.
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:25049233,
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:25049233};
CC Note=Binds 1 [2Fe-2S] cluster per subunit. {ECO:0000255|PROSITE-
CC ProRule:PRU00628, ECO:0000269|PubMed:25049233};
CC -!- COFACTOR:
CC Name=Fe cation; Xref=ChEBI:CHEBI:24875;
CC Evidence={ECO:0000269|PubMed:25049233};
CC Note=Binds 1 Fe cation. {ECO:0000269|PubMed:25049233};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.5 uM for 1,4-BNC as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC KM=0.5 uM for testosterone as substrate (at pH 7.0 and 22 degrees
CC Celsius) {ECO:0000269|PubMed:25049233};
CC KM=0.8 uM for AD as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC KM=1.2 uM for 4-BNC as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC KM=6.1 uM for 5alpha-androstan-3,17-dione as substrate (at pH 7.0 and
CC 22 degrees Celsius) {ECO:0000269|PubMed:25049233};
CC KM=500 uM for 1,4-BNC-CoA as substrate (at pH 7.0 and 22 degrees
CC Celsius) {ECO:0000269|PubMed:25049233};
CC Note=kcat is 1.7 sec(-1) for 1,4-BNC-CoA as substrate (at pH 7.0 and
CC 22 degrees Celsius). kcat is 0.8 sec(-1) for 5alpha-androstan-3,17-
CC dione, AD and testosterone as substrates (at pH 7.0 and 22 degrees
CC Celsius). kcat is 0.7 sec(-1) for ADD as substrate (at pH 7.0 and 22
CC degrees Celsius). kcat is 0.6 sec(-1) for 4-BNC as substrate (at pH
CC 7.0 and 22 degrees Celsius). kcat is 0.5 sec(-1) for 1,4-BNC as
CC substrate (at pH 7.0 and 22 degrees Celsius).
CC {ECO:0000269|PubMed:25049233};
CC -!- PATHWAY: Steroid metabolism; cholesterol degradation.
CC {ECO:0000305|PubMed:25049233}.
CC -!- SUBUNIT: Homotrimer. The two-component system 3-ketosteroid-9-alpha-
CC monooxygenase is composed of an oxygenase component KshA and a
CC reductase component KshB. {ECO:0000269|PubMed:25049233}.
CC -!- INDUCTION: By cholesterol, progesterone, AD and cholic acid.
CC {ECO:0000269|PubMed:21642460}.
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DR EMBL; HQ425877; ADY18328.1; -; Genomic_DNA.
DR PDB; 4QDC; X-ray; 1.90 A; A=1-390.
DR PDB; 4QDD; X-ray; 2.60 A; A=1-390.
DR PDB; 4QDF; X-ray; 2.43 A; A=1-390.
DR PDBsum; 4QDC; -.
DR PDBsum; 4QDD; -.
DR PDBsum; 4QDF; -.
DR AlphaFoldDB; F1CMY8; -.
DR SMR; F1CMY8; -.
DR BRENDA; 1.14.15.30; 5395.
DR UniPathway; UPA01058; -.
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:0006707; P:cholesterol catabolic process; NAS:UniProtKB.
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; 3D-structure; Cholesterol metabolism; Iron; Iron-sulfur;
KW Lipid degradation; Lipid metabolism; Metal-binding; Oxidoreductase;
KW Steroid metabolism; Sterol metabolism.
FT CHAIN 1..390
FT /note="3-ketosteroid-9-alpha-monooxygenase, oxygenase
FT component"
FT /id="PRO_0000438403"
FT DOMAIN 32..134
FT /note="Rieske"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 73
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628,
FT ECO:0000269|PubMed:25049233"
FT BINDING 75
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628,
FT ECO:0000269|PubMed:25049233"
FT BINDING 92
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628,
FT ECO:0000269|PubMed:25049233"
FT BINDING 95
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628,
FT ECO:0000269|PubMed:25049233"
FT BINDING 181
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25049233"
FT BINDING 187
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25049233"
FT BINDING 192
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25049233"
FT BINDING 311
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25049233"
FT STRAND 31..37
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 38..41
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 43..45
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 47..51
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 54..60
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 62..64
FT /evidence="ECO:0007829|PDB:4QDD"
FT STRAND 66..72
FT /evidence="ECO:0007829|PDB:4QDC"
FT TURN 74..76
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 80..82
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 83..86
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 89..91
FT /evidence="ECO:0007829|PDB:4QDC"
FT TURN 93..95
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 98..100
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 103..107
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 111..113
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 125..128
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 131..136
FT /evidence="ECO:0007829|PDB:4QDC"
FT TURN 153..156
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 164..172
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 176..179
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 180..183
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 185..190
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 194..204
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 207..216
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 223..225
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 232..253
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 256..270
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 273..283
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 286..288
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 290..317
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 333..341
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 342..344
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 347..349
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 352..355
FT /evidence="ECO:0007829|PDB:4QDC"
FT STRAND 358..362
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 365..377
FT /evidence="ECO:0007829|PDB:4QDC"
FT HELIX 380..382
FT /evidence="ECO:0007829|PDB:4QDC"
SQ SEQUENCE 390 AA; 44249 MW; 46E13D2C9B6F7025 CRC64;
MSIDTARSGS DDDVEIREIQ AAAAPTRFAR GWHCLGLLRD FQDGKPHSIE AFGTKLVVFA
DSKGQLNVLD AYCRHMGGDL SRGEVKGDSI ACPFHDWRWN GKGKCTDIPY ARRVPPIAKT
RAWTTLERNG QLYVWNDPQG NPPPEDVTIP EIAGYGTDEW TDWSWKSLRI KGSHCREIVD
NVVDMAHFFY IHYSFPRYFK NVFEGHTATQ YMHSTGREDV ISGTNYDDPN AELRSEATYF
GPSYMIDWLE SDANGQTIET ILINCHYPVS NNEFVLQYGA IVKKLPGVSD EIAAGMAEQF
AEGVQLGFEQ DVEIWKNKAP IDNPLLSEED GPVYQLRRWY QQFYVDVEDI TEDMTKRFEF
EIDTTRAVAS WQKEVAENLA KQAEGSTATP
