CPXM_PRIMG
ID CPXM_PRIMG Reviewed; 410 AA.
AC Q06069;
DT 01-FEB-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1995, sequence version 1.
DT 03-AUG-2022, entry version 101.
DE RecName: Full=Cytochrome P450(MEG);
DE EC=1.14.99.-;
DE AltName: Full=Steroid 15-beta-hydroxylase;
DE EC=1.14.15.8;
DE AltName: Full=Steroid 15-beta-monooxygenase;
GN Name=cyp106A2;
OS Priestia megaterium (Bacillus megaterium).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Priestia.
OX NCBI_TaxID=1404;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 13368 / 41;
RX PubMed=8232201; DOI=10.1007/bf00280214;
RA Rauschenbach R., Isernhagen M., Noeske-Jungblut C., Boidol W., Siewert G.;
RT "Cloning sequencing and expression of the gene for cytochrome P450meg, the
RT steroid-15 beta-monooxygenase from Bacillus megaterium ATCC 13368.";
RL Mol. Gen. Genet. 241:170-176(1993).
RN [2]
RP CHARACTERIZATION.
RC STRAIN=ATCC 13368 / 41;
RX PubMed=109432; DOI=10.1016/s0021-9258(18)50589-5;
RA Berg A., Ingelman-Sundberg M., Gustafsson M.;
RT "Purification and characterization of cytochrome P-450meg.";
RL J. Biol. Chem. 254:5264-5271(1979).
RN [3]
RP CHARACTERIZATION.
RC STRAIN=ATCC 13368 / 41;
RX PubMed=6797409; DOI=10.1042/bj1960781;
RA Berg A., Rafter J.J.;
RT "Studies on the substrate specificity and inducibility of cytochrome P-
RT 450meg.";
RL Biochem. J. 196:781-786(1981).
RN [4]
RP CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=15178459; DOI=10.1016/j.bbrc.2004.05.037;
RA Lisurek M., Kang M.J., Hartmann R.W., Bernhardt R.;
RT "Identification of monohydroxy progesterones produced by CYP106A2 using
RT comparative HPLC and electrospray ionisation collision-induced dissociation
RT mass spectrometry.";
RL Biochem. Biophys. Res. Commun. 319:677-682(2004).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF SER-394; ALA-395; THR-396; GLY-397 AND GLN-398.
RX PubMed=18481342; DOI=10.1002/cbic.200700670;
RA Lisurek M., Simgen B., Antes I., Bernhardt R.;
RT "Theoretical and experimental evaluation of a CYP106A2 low homology model
RT and production of mutants with changed activity and selectivity of
RT hydroxylation.";
RL ChemBioChem 9:1439-1449(2008).
CC -!- FUNCTION: Has the capacity to hydroxylate certain steroids in the 15-
CC beta position. Also hydroxylates progesterone in the 11-alpha and 9-
CC beta position. {ECO:0000269|PubMed:18481342}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 H(+) + O2 + progesterone + reduced 2[4Fe-4S]-[ferredoxin] =
CC 15beta-hydroxyprogesterone + H2O + oxidized 2[4Fe-4S]-[ferredoxin];
CC Xref=Rhea:RHEA:27337, Rhea:RHEA-COMP:10002, Rhea:RHEA-COMP:10004,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17026, ChEBI:CHEBI:33722, ChEBI:CHEBI:33723,
CC ChEBI:CHEBI:59418; EC=1.14.15.8;
CC Evidence={ECO:0000269|PubMed:15178459, ECO:0000269|PubMed:18481342};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413; Evidence={ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=251.4 uM for progesterone for the 15-beta hydroxylation
CC {ECO:0000269|PubMed:15178459, ECO:0000269|PubMed:18481342};
CC KM=77.5 uM for progesterone for the 6-beta hydroxylation
CC {ECO:0000269|PubMed:15178459, ECO:0000269|PubMed:18481342};
CC KM=140.3 uM for progesterone for the 11-alpha hydroxylation
CC {ECO:0000269|PubMed:15178459, ECO:0000269|PubMed:18481342};
CC KM=155.5 uM for progesterone for the 9-alpha hydroxylation
CC {ECO:0000269|PubMed:15178459, ECO:0000269|PubMed:18481342};
CC KM=1.5 uM for deoxycorticosterone {ECO:0000269|PubMed:15178459,
CC ECO:0000269|PubMed:18481342};
CC Vmax=337 nmol/min/mg enzyme for the 15-beta hydroxylation toward
CC progesterone {ECO:0000269|PubMed:15178459,
CC ECO:0000269|PubMed:18481342};
CC Vmax=22.3 nmol/min/mg enzyme for the 6-beta hydroxylation toward
CC progesterone {ECO:0000269|PubMed:15178459,
CC ECO:0000269|PubMed:18481342};
CC Vmax=17.5 nmol/min/mg enzyme for the 11-alpha hydroxylation toward
CC progesterone {ECO:0000269|PubMed:15178459,
CC ECO:0000269|PubMed:18481342};
CC Vmax=6.5 nmol/min/mg enzyme for the 6-alpha hydroxylation toward
CC progesterone {ECO:0000269|PubMed:15178459,
CC ECO:0000269|PubMed:18481342};
CC Vmax=246 nmol/min/mg enzyme toward deoxycorticosterone
CC {ECO:0000269|PubMed:15178459, ECO:0000269|PubMed:18481342};
CC Note=With deoxycorticosterone as substrate only 15-beta hydroxylation
CC is observed.;
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; Z21972; CAA79985.1; -; Genomic_DNA.
DR PIR; S39924; S39924.
DR PDB; 4YT3; X-ray; 1.80 A; A/B=1-410.
DR PDB; 5IKI; X-ray; 2.40 A; A/B=1-410.
DR PDB; 5XNT; X-ray; 2.70 A; A=1-409.
DR PDBsum; 4YT3; -.
DR PDBsum; 5IKI; -.
DR PDBsum; 5XNT; -.
DR AlphaFoldDB; Q06069; -.
DR SMR; Q06069; -.
DR KEGG; ag:CAA79985; -.
DR BioCyc; MetaCyc:MON-15437; -.
DR BRENDA; 1.14.15.8; 656.
DR BRENDA; 1.14.99.14; 656.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR002397; Cyt_P450_B.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00359; BP450.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Heme; Iron; Metal-binding; Monooxygenase;
KW Oxidoreductase.
FT CHAIN 1..410
FT /note="Cytochrome P450(MEG)"
FT /id="PRO_0000052219"
FT BINDING 355
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250"
FT MUTAGEN 394
FT /note="S->I: 30% of 15-beta hydroxylation activity (for
FT progesterone)."
FT /evidence="ECO:0000269|PubMed:18481342"
FT MUTAGEN 395
FT /note="A->L: 40% of 15-beta hydroxylation activity (for
FT progesterone)."
FT /evidence="ECO:0000269|PubMed:18481342"
FT MUTAGEN 396
FT /note="T->R: Loss of 15-beta hydroxylation activity (for
FT progesterone and deoxycorticosterone)."
FT /evidence="ECO:0000269|PubMed:18481342"
FT MUTAGEN 397
FT /note="G->P: Loss of 15-beta hydroxylation activity (for
FT progesterone)."
FT /evidence="ECO:0000269|PubMed:18481342"
FT MUTAGEN 398
FT /note="Q->S: 30% of 15-beta hydroxylation activity (for
FT progesterone)."
FT /evidence="ECO:0000269|PubMed:18481342"
FT HELIX 7..11
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 16..20
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 23..32
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 34..38
FT /evidence="ECO:0007829|PDB:4YT3"
FT TURN 39..42
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 43..46
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 49..57
FT /evidence="ECO:0007829|PDB:4YT3"
FT TURN 59..61
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 62..65
FT /evidence="ECO:0007829|PDB:4YT3"
FT TURN 78..80
FT /evidence="ECO:0007829|PDB:5IKI"
FT HELIX 84..86
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 88..90
FT /evidence="ECO:0007829|PDB:5XNT"
FT HELIX 95..104
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 109..128
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 132..134
FT /evidence="ECO:0007829|PDB:5IKI"
FT STRAND 135..137
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 138..141
FT /evidence="ECO:0007829|PDB:4YT3"
FT TURN 142..144
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 145..155
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 159..161
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 162..173
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 184..208
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 214..220
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 230..261
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 266..272
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 274..276
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 277..287
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 293..298
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 306..309
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 314..318
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 319..322
FT /evidence="ECO:0007829|PDB:4YT3"
FT TURN 326..328
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 329..331
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 342..344
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 351..353
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 358..375
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 377..381
FT /evidence="ECO:0007829|PDB:4YT3"
FT HELIX 387..390
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 391..394
FT /evidence="ECO:0007829|PDB:5XNT"
FT STRAND 399..401
FT /evidence="ECO:0007829|PDB:4YT3"
FT STRAND 404..408
FT /evidence="ECO:0007829|PDB:4YT3"
SQ SEQUENCE 410 AA; 46956 MW; 9FDD9CF2EE0F810B CRC64;
MKEVIAVKEI TRFKTRTEEF SPYAWCKRML ENDPVSYHEG TDTWNVFKYE DVKRVLSDYK
HFSSVRKRTT ISVGTDSEEG SVPEKIQITE SDPPDHRKRR SLLAAAFTPR SLQNWEPRIQ
EIADELIGQM DGGTEIDIVA SLASPLPIIV MADLMGVPSK DRLLFKKWVD TLFLPFDREK
QEEVDKLKQV AAKEYYQYLY PIVVQKRLNP ADDIISDLLK SEVDGEMFTD DEVVRTTMLI
LGAGVETTSH LLANSFYSLL YDDKEVYQEL HENLDLVPQA VEEMLRFRFN LIKLDRTVKE
DNDLLGVELK EGDSVVVWMS AANMDEEMFE DPFTLNIHRP NNKKHLTFGN GPHFCLGAPL
ARLEAKIALT AFLKKFKHIE AVPSFQLEEN LTDSATGQTL TSLPLKASRM
