KSHA1_RHORH
ID KSHA1_RHORH Reviewed; 394 AA.
AC F1CMX0;
DT 30-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 03-MAY-2011, sequence version 1.
DT 03-AUG-2022, entry version 40.
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=kshA1 {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 (2.43 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG;
RP IRON ION AND IRON-SULFUR (2FE-2S), FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, SUBUNIT, AND SUBSTRATE
RP 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: May be involved in the degradation of cholic acid, a steroid
CC acid found predominantly in the bile (PubMed:21642460). In vitro,
CC catalyzes the introduction of a 9alpha-hydroxyl moiety into the ring B
CC of 3-ketosteroid substrates such as 1,4-androstadiene-3,17-dione (ADD),
CC 4-androstene-3,17-dione (AD), 4-androstene-17beta-ol-3-one
CC (testosterone), 4-pregnene-3,20-dione (progesterone), 3-oxo-23,24-
CC bisnorcholesta-4-en-22-oate (4-BNC), 23,24-bisnorcholesta-4-ene-22-
CC oate, 3-oxo-23,24-bisnorcholaesta-1,4-dien-22-oate (1,4-BNC), 23,24-
CC bisnorcholesta-1,4-diene-22-oate and 3-oxo-23,24-bisnorcholesta-1,4-
CC dien-22-oyl-coenzyme A thioester (1,4-BNC-CoA) (PubMed:21642460,
CC PubMed:25049233). KshA1 has the highest specificity for steroids
CC possessing an isopropionyl side chain at C17 (PubMed:25049233).
CC {ECO:0000269|PubMed:21642460, 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=1 uM for 1,4-BNC as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC KM=2 uM for 1,4-BNC-CoA as substrate (at pH 7.0 and 22 degrees
CC Celsius) {ECO:0000269|PubMed:25049233};
CC KM=2.2 uM for 4-BNC as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC KM=50 uM for ADD as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC KM=110 uM for testosterone as substrate (at pH 7.0 and 22 degrees
CC Celsius) {ECO:0000269|PubMed:25049233};
CC KM=400 uM for AD as substrate (at pH 7.0 and 22 degrees Celsius)
CC {ECO:0000269|PubMed:25049233};
CC Note=kcat is 2.6 sec(-1) for 4-BNC as substrate (at pH 7.0 and 22
CC degrees Celsius). kcat is 1.4 sec(-1) for testosterone as substrate
CC (at pH 7.0 and 22 degrees Celsius). kcat is 1.3 sec(-1) for ADD as
CC substrate (at pH 7.0 and 22 degrees Celsius). kcat is 1.1 sec(-1) for
CC 1,4-BNC-CoA as substrate (at pH 7.0 and 22 degrees Celsius). kcat is
CC 0.9 sec(-1) for 1,4-BNC as substrate (at pH 7.0 and 22 degrees
CC Celsius). kcat is 0.5 sec(-1) for AD as substrate (at pH 7.0 and 22
CC degrees Celsius). {ECO:0000269|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 cholic acid. {ECO:0000269|PubMed:21642460}.
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DR EMBL; HQ425873; ADY18310.1; -; Genomic_DNA.
DR RefSeq; WP_059382524.1; NZ_LT906450.1.
DR PDB; 4QDF; X-ray; 2.43 A; B=1-394.
DR PDBsum; 4QDF; -.
DR AlphaFoldDB; F1CMX0; -.
DR SMR; F1CMX0; -.
DR PRIDE; F1CMX0; -.
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; 3D-structure; Iron; Iron-sulfur; Lipid degradation;
KW Lipid metabolism; Metal-binding; Oxidoreductase; Steroid metabolism.
FT CHAIN 1..394
FT /note="3-ketosteroid-9-alpha-monooxygenase, oxygenase
FT component"
FT /id="PRO_0000438399"
FT DOMAIN 27..129
FT /note="Rieske"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 68
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 70
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 87
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 90
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 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:0000269|PubMed:25049233"
FT BINDING 245
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25049233"
FT BINDING 305
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25049233"
FT STRAND 26..32
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 33..36
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 38..40
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 42..46
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 49..55
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 61..67
FT /evidence="ECO:0007829|PDB:4QDF"
FT TURN 69..71
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 75..77
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 78..81
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 84..86
FT /evidence="ECO:0007829|PDB:4QDF"
FT TURN 88..90
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 98..102
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 120..123
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 126..131
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 148..151
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 159..167
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 170..173
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 174..177
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 181..184
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 188..198
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 201..210
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 226..246
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 251..264
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 267..277
FT /evidence="ECO:0007829|PDB:4QDF"
FT TURN 284..286
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 287..310
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 327..334
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 335..338
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 341..343
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 346..349
FT /evidence="ECO:0007829|PDB:4QDF"
FT STRAND 352..356
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 359..375
FT /evidence="ECO:0007829|PDB:4QDF"
FT HELIX 376..378
FT /evidence="ECO:0007829|PDB:4QDF"
FT TURN 379..383
FT /evidence="ECO:0007829|PDB:4QDF"
SQ SEQUENCE 394 AA; 45020 MW; 2E7CBC44AB112CFE CRC64;
MSLGTSEQSE IREIVAGSAP ARFARGWHCL GLAKDFKDGK PHSVHAFGTK LVVWADSNDE
IRILDAYCRH MGGDLSQGTV KGDEIACPFH DWRWGGNGRC KNIPYARRVP PIAKTRAWHT
LDQDGLLFVW HDPQGNPPPA DVTIPRIAGA TSDEWTDWVW YTTEVDTNCR EIIDNIVDMA
HFFYVHYSFP VYFKNVFEGH VASQFMRGQA REDTRPHANG QPKMIGSRSD ASYFGPSFMI
DDLVYEYEGY DVESVLINCH YPVSQDKFVL MYGMIVKKSD RLEGEKALQT AQQFGNFIAK
GFEQDIEIWR NKTRIDNPLL CEEDGPVYQL RRWYEQFYVD VEDVAPEMTD RFEFEMDTTR
PVAAWMKEVE ANIARKAALD TETRSAPEQS TTAG
