KSHB_MYCTU
ID KSHB_MYCTU Reviewed; 358 AA.
AC P9WJ93; L0TD71; P96853; Q7D594;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 46.
DE RecName: Full=3-ketosteroid-9-alpha-monooxygenase, ferredoxin reductase component {ECO:0000303|PubMed:19234303};
DE AltName: Full=3-ketosteroid-9-alpha-hydroxylase, ferredoxin reductase component {ECO:0000303|PubMed:19234303};
DE Short=KSH {ECO:0000303|PubMed:19234303};
DE AltName: Full=Androsta-1,4-diene-3,17-dione 9-alpha-hydroxylase {ECO:0000303|PubMed:19234303};
DE EC=1.14.15.30 {ECO:0000269|PubMed:19234303, ECO:0000269|PubMed:21987574};
DE AltName: Full=Rieske-type oxygenase {ECO:0000303|PubMed:19234303};
DE Short=RO {ECO:0000303|PubMed:19234303};
GN Name=hmp; Synonyms=kshB; OrderedLocusNames=Rv3571;
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2]
RP INDUCTION.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=17635188; DOI=10.1111/j.1365-2958.2007.05827.x;
RA Kendall S.L., Withers M., Soffair C.N., Moreland N.J., Gurcha S.,
RA Sidders B., Frita R., Ten Bokum A., Besra G.S., Lott J.S., Stoker N.G.;
RT "A highly conserved transcriptional repressor controls a large regulon
RT involved in lipid degradation in Mycobacterium smegmatis and Mycobacterium
RT tuberculosis.";
RL Mol. Microbiol. 65:684-699(2007).
RN [3]
RP FUNCTION AS A 3-KETOSTEROID-9-ALPHA-HYDROXYLASE, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, PATHWAY, AND SUBSTRATE
RP SPECIFICITY.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=19234303; DOI=10.1074/jbc.m900719200;
RA Capyk J.K., D'Angelo I., Strynadka N.C., Eltis L.D.;
RT "Characterization of 3-ketosteroid 9{alpha}-hydroxylase, a Rieske oxygenase
RT in the cholesterol degradation pathway of Mycobacterium tuberculosis.";
RL J. Biol. Chem. 284:9937-9946(2009).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, AND SUBUNIT.
RX PubMed=21987574; DOI=10.1074/jbc.m111.289975;
RA Capyk J.K., Casabon I., Gruninger R., Strynadka N.C., Eltis L.D.;
RT "Activity of 3-ketosteroid 9alpha-hydroxylase (KshAB) indicates cholesterol
RT side chain and ring degradation occur simultaneously in Mycobacterium
RT tuberculosis.";
RL J. Biol. Chem. 286:40717-40724(2011).
RN [5]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
CC -!- FUNCTION: Involved in the degradation of cholesterol. Catalyzes the
CC introduction of a 9a-hydroxyl moiety into 1,4-androstadiene-3,17-dione
CC (ADD) to yield the 9alpha-hydroxy-1,4-androstadiene-3,17-dione (9OHADD)
CC intermediate which spontaneously form 3-hydroxy-9,10-seconandrost-
CC 1,3,5(10)-triene-9,17-dione (HSA) via the meta-cleavage of ring B with
CC concomitant aromatization of ring A. KSH is also able to use 4-
CC androstene-3,17-dione (AD), 3-oxo-23,24-bisnorcholesta-4-en-22-oate (4-
CC BNC), 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate (1,4-BNC), 3-oxo-
CC 23,24-bisnorcholesta-4-en-22-oyl-coenzyme A thioester (4-BNC-CoA) and
CC 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-coenzyme A thioester (1,4-
CC BNC-CoA) as substrates. {ECO:0000269|PubMed:19234303,
CC ECO:0000269|PubMed:21987574}.
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:19234303,
CC ECO:0000269|PubMed:21987574};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:19234303};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:19234303};
CC -!- COFACTOR:
CC Name=[2Fe-2S] cluster; Xref=ChEBI:CHEBI:190135;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU00465,
CC ECO:0000269|PubMed:19234303};
CC Note=Binds 1 2Fe-2S cluster. {ECO:0000255|PROSITE-ProRule:PRU00465,
CC ECO:0000269|PubMed:19234303};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3 uM for 4-BNC (at pH 7 and at 22 degrees Celsius)
CC {ECO:0000269|PubMed:21987574};
CC KM=6.8 uM for 4-BNC-CoA (at pH 7 and at 22 degrees Celsius)
CC {ECO:0000269|PubMed:21987574};
CC KM=17 uM for 1,4-BNC-CoA (at pH 7 and at 22 degrees Celsius)
CC {ECO:0000269|PubMed:21987574};
CC KM=24 uM for AD (at pH 7 and at 25 degrees Celsius)
CC {ECO:0000269|PubMed:19234303};
CC KM=70 uM for 1,4-BNC (at pH 7 and at 22 degrees Celsius)
CC {ECO:0000269|PubMed:21987574};
CC KM=110 uM for ADD (at pH 7 and at 25 degrees Celsius)
CC {ECO:0000269|PubMed:19234303};
CC Note=kcat is 2.7 sec(-1) for 1,4-BNC-CoA as substrate (at pH 7 and at
CC 22 degrees Celsius) (PubMed:21987574). kcat is 0.8 sec(-1) for ADD as
CC substrate (at pH 7 and at 25 degrees Celsius) (PubMed:19234303). kcat
CC is 0.61 sec(-1) for 4-BNC-CoA as substrate (at pH 7 and at 22 degrees
CC Celsius) (PubMed:21987574). kcat is 0.25 sec(-1) for 1,4-BNC as
CC substrate (at pH 7 and at 22 degrees Celsius) (PubMed:21987574). kcat
CC is 0.08 sec(-1) for 4-BNC as substrate (at pH 7 and at 22 degrees
CC Celsius) (PubMed:21987574). kcat is 0.07 sec(-1) for AD as substrate
CC (at pH 7 and at 25 degrees Celsius) (PubMed:19234303).
CC {ECO:0000269|PubMed:19234303, ECO:0000269|PubMed:21987574};
CC -!- PATHWAY: Lipid metabolism; steroid biosynthesis.
CC {ECO:0000305|PubMed:19234303}.
CC -!- SUBUNIT: Monomer. 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:21987574}.
CC -!- INDUCTION: Induced by KstR. {ECO:0000269|PubMed:17635188}.
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DR EMBL; AL123456; CCP46394.1; -; Genomic_DNA.
DR PIR; A70606; A70606.
DR RefSeq; NP_218088.1; NC_000962.3.
DR RefSeq; WP_003900102.1; NZ_NVQJ01000014.1.
DR AlphaFoldDB; P9WJ93; -.
DR SMR; P9WJ93; -.
DR STRING; 83332.Rv3571; -.
DR PaxDb; P9WJ93; -.
DR DNASU; 887315; -.
DR GeneID; 887315; -.
DR KEGG; mtu:Rv3571; -.
DR TubercuList; Rv3571; -.
DR eggNOG; COG1018; Bacteria.
DR OMA; ILACQSH; -.
DR PhylomeDB; P9WJ93; -.
DR BioCyc; MetaCyc:G185E-7849-MON; -.
DR BRENDA; 1.14.15.30; 3445.
DR UniPathway; UPA00062; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0051537; F:2 iron, 2 sulfur cluster binding; IDA:MTBBASE.
DR GO; GO:0036200; F:3-ketosteroid 9-alpha-monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0071949; F:FAD binding; IDA:MTBBASE.
DR GO; GO:0047086; F:ketosteroid monooxygenase activity; IDA:MTBBASE.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0006707; P:cholesterol catabolic process; IDA:MTBBASE.
DR GO; GO:0008203; P:cholesterol metabolic process; IDA:MTBBASE.
DR GO; GO:0001666; P:response to hypoxia; IEP:MTBBASE.
DR GO; GO:0051409; P:response to nitrosative stress; IEP:MTBBASE.
DR GO; GO:0006694; P:steroid biosynthetic process; IEA:UniProtKB-UniPathway.
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 Reference proteome; Steroid metabolism; Sterol metabolism; Virulence.
FT CHAIN 1..358
FT /note="3-ketosteroid-9-alpha-monooxygenase, ferredoxin
FT reductase component"
FT /id="PRO_0000404100"
FT DOMAIN 12..124
FT /note="FAD-binding FR-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT DOMAIN 269..358
FT /note="2Fe-2S ferredoxin-type"
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 310
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT BINDING 313
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
FT BINDING 343
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00465"
SQ SEQUENCE 358 AA; 38265 MW; 182A799D00ADC8B2 CRC64;
MTEAIGDEPL GDHVLELQIA EVVDETDEAR SLVFAVPDGS DDPEIPPRRL RYAPGQFLTL
RVPSERTGSV ARCYSLCSSP YTDDALAVTV KRTADGYASN WLCDHAQVGM RIHVLAPSGN
FVPTTLDADF LLLAAGSGIT PIMSICKSAL AEGGGQVTLL YANRDDRSVI FGDALRELAA
KYPDRLTVLH WLESLQGLPS ASALAKLVAP YTDRPVFICG PGPFMQAARD ALAALKVPAQ
QVHIEVFKSL ESDPFAAVKV DDSGDEAPAT AVVELDGQTH TVSWPRTAKL LDVLLAAGLD
APFSCREGHC GACACTLRAG KVNMGVNDVL EQQDLDEGLI LACQSRPESD SVEVTYDE
