CPXE_STRGO
ID CPXE_STRGO Reviewed; 406 AA.
AC P18326;
DT 01-NOV-1990, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 2.
DT 03-AUG-2022, entry version 105.
DE RecName: Full=Vitamin D3 dihydroxylase {ECO:0000305};
DE EC=1.14.15.22 {ECO:0000269|PubMed:15207715, ECO:0000269|PubMed:18314962, ECO:0000269|PubMed:18937506};
DE AltName: Full=CYP105A1 {ECO:0000303|PubMed:2345149};
DE AltName: Full=Cytochrome P450-CVA1 {ECO:0000303|PubMed:2345149};
DE AltName: Full=Cytochrome P450-SU1 {ECO:0000303|PubMed:2345149};
DE AltName: Full=Vitamin D3 hydroxylase {ECO:0000303|PubMed:18937506};
DE Short=VD3 hydroxylase {ECO:0000303|PubMed:18937506};
GN Name=cyp105A1 {ECO:0000303|PubMed:2345149};
GN Synonyms=suaC {ECO:0000303|PubMed:2345149};
OS Streptomyces griseolus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1909;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-32, AND INDUCTION.
RC STRAIN=ATCC 11796 / DSM 40854;
RX PubMed=2345149; DOI=10.1128/jb.172.6.3335-3345.1990;
RA Omer C.A., Lenstra R., Litle P.J., Dean C., Tepperman J.M., Leto K.J.,
RA Romesser J.A., O'Keefe D.P.;
RT "Genes for two herbicide-inducible cytochromes P-450 from Streptomyces
RT griseolus.";
RL J. Bacteriol. 172:3335-3345(1990).
RN [2]
RP PROTEIN SEQUENCE OF 2-11, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBCELLULAR LOCATION, COFACTOR, AND SUBSTRATE SPECIFICITY.
RX PubMed=15207715; DOI=10.1016/j.bbrc.2004.05.140;
RA Sawada N., Sakaki T., Yoneda S., Kusudo T., Shinkyo R., Ohta M., Inouye K.;
RT "Conversion of vitamin D3 to 1alpha,25-dihydroxyvitamin D3 by Streptomyces
RT griseolus cytochrome P450SU-1.";
RL Biochem. Biophys. Res. Commun. 320:156-164(2004).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF WILD-TYPE AND MUTANT ALA-84 IN
RP COMPLEX WITH HEME AND CALCITRIOL, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ARG-73; ARG-84; VAL-88;
RP LEU-180; VAL-181; ARG-193 AND ILE-293, AND COFACTOR.
RX PubMed=18314962; DOI=10.1021/bi7023767;
RA Sugimoto H., Shinkyo R., Hayashi K., Yoneda S., Yamada M., Kamakura M.,
RA Ikushiro S., Shiro Y., Sakaki T.;
RT "Crystal structure of CYP105A1 (P450SU-1) in complex with 1alpha,25-
RT dihydroxyvitamin D3.";
RL Biochemistry 47:4017-4027(2008).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF MUTANT PHE-84 AND DOUBLE MUTANT
RP ALA-73/ALA-84 IN COMPLEX WITH HEME AND CALCITRIOL, FUNCTION, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ARG-73 AND ARG-84,
RP AND COFACTOR.
RX PubMed=18937506; DOI=10.1021/bi801222d;
RA Hayashi K., Sugimoto H., Shinkyo R., Yamada M., Ikeda S., Ikushiro S.,
RA Kamakura M., Shiro Y., Sakaki T.;
RT "Structure-based design of a highly active vitamin D hydroxylase from
RT Streptomyces griseolus CYP105A1.";
RL Biochemistry 47:11964-11972(2008).
CC -!- FUNCTION: Involved in the metabolism of vitamin D3 (calciol) and of a
CC number of sulfonylurea herbicides. Catalyzes the two-step hydroxylation
CC (25- and 1-alpha-hydroxylation) of vitamin D3 (VD3) to yield its active
CC form 1-alpha,25-dihydroxyvitamin D3 (calcitriol). The first step is the
CC hydroxylation of the C-25 position of VD3 to produce 25-hydroxyvitamin
CC D3 (calcidiol). The second reaction is the hydroxylation of the C1-
CC alpha-position of calcidiol to produce calcitriol. It can also
CC hydroxylate vitamin D2. {ECO:0000269|PubMed:15207715,
CC ECO:0000269|PubMed:18314962, ECO:0000269|PubMed:18937506}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=calciol + 2 H(+) + O2 + 2 reduced [2Fe-2S]-[ferredoxin] =
CC calcidiol + H2O + 2 oxidized [2Fe-2S]-[ferredoxin];
CC Xref=Rhea:RHEA:50696, Rhea:RHEA-COMP:10000, Rhea:RHEA-COMP:10001,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17933, ChEBI:CHEBI:28940, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738; EC=1.14.15.22;
CC Evidence={ECO:0000269|PubMed:15207715, ECO:0000269|PubMed:18314962,
CC ECO:0000269|PubMed:18937506};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=calcidiol + 2 H(+) + O2 + 2 reduced [2Fe-2S]-[ferredoxin] =
CC calcitriol + H2O + 2 oxidized [2Fe-2S]-[ferredoxin];
CC Xref=Rhea:RHEA:50700, Rhea:RHEA-COMP:10000, Rhea:RHEA-COMP:10001,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17823, ChEBI:CHEBI:17933, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738; EC=1.14.15.22;
CC Evidence={ECO:0000269|PubMed:15207715, ECO:0000269|PubMed:18314962,
CC ECO:0000269|PubMed:18937506};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000269|PubMed:15207715, ECO:0000269|PubMed:18314962,
CC ECO:0000269|PubMed:18937506};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.54 uM for vitamin D3 (at pH 7.4 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:15207715};
CC KM=0.59 uM for vitamin D2 (at pH 7.4 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:15207715};
CC KM=0.91 uM for 25-hydroxyvitamin D3 (1-alpha-hydroxylation)(at pH 7.4
CC and 30 degrees Celsius) {ECO:0000269|PubMed:15207715};
CC KM=4.4 uM for 25-hydroxyvitamin D3 (1-alpha-hydroxylation)
CC {ECO:0000269|PubMed:18937506};
CC KM=5 uM for 25-hydroxyvitamin D3 (1-alpha-hydroxylation)
CC {ECO:0000269|PubMed:18314962};
CC KM=9.4 uM for 1-alpha-hydroxyvitamin D3 (25-alpha-hydroxylation)
CC {ECO:0000269|PubMed:18314962};
CC KM=10.1 uM for 1-alpha-hydroxyvitamin D3 (25-alpha-hydroxylation)
CC {ECO:0000269|PubMed:18937506};
CC Vmax=8.4 umol/min/mg enzyme with 25-hydroxyvitamin D3 as substrate
CC (1-alpha-hydroxylation) {ECO:0000269|PubMed:18314962};
CC Vmax=3.8 umol/min/mg enzyme with as 1-alpha-hydroxyvitamin D3
CC substrate (25-alpha-hydroxylation) {ECO:0000269|PubMed:18314962};
CC Vmax=3.6 mmol/min/mg enzyme with 25-hydroxyvitamin D3 as substrate
CC (1-alpha-hydroxylation)(at pH 7.4 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:15207715};
CC Vmax=16 mmol/min/mg enzyme with vitamin D3 as substrate (at pH 7.4
CC and 30 degrees Celsius) {ECO:0000269|PubMed:15207715};
CC Vmax=84 mmol/min/mg enzyme with vitamin D2 as substrate (at pH 7.4
CC and 30 degrees Celsius) {ECO:0000269|PubMed:15207715};
CC Note=kcat is 0.0076 min(-1) for hydroxylase activity with 1-alpha-
CC hydroxyvitamin D3 as substrate. kcat is 0.0026 min(-1) for
CC hydroxylase activity with 25-hydroxyvitamin D3 as substrate.
CC {ECO:0000269|PubMed:18937506};
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:15207715}.
CC -!- INDUCTION: By herbicides. {ECO:0000269|PubMed:2345149}.
CC -!- MISCELLANEOUS: Two metabolic pathways exist. One is a conversion of VD3
CC to 25-hydroxyvitamin D3 and then to 1-alpha,25-dihydroxyvitamin D3 (25-
CC pathway), while the other one is a pathway via 1-alpha-hydroxyvitamin
CC D3 as an intermediate (1-alpha-pathway). Analysis show that the amount
CC of 25-hydroxyvitamin D3 is predominant as the product of this reaction,
CC while very small amounts of 1-alpha-hydroxyvitamin D3 and 1-alpha,25-
CC dihydroxyvitamin D3 are obtained, suggesting that the major pathway is
CC the 25-pathway. {ECO:0000269|PubMed:18937506}.
CC -!- MISCELLANEOUS: The hydrophobicity and the volume of the side chain at
CC position 84 are critical for the activity.
CC {ECO:0000269|PubMed:18937506}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; M32238; AAA26823.1; -; Genomic_DNA.
DR PIR; A35401; A35401.
DR PDB; 2ZBX; X-ray; 1.50 A; A=1-406.
DR PDB; 2ZBY; X-ray; 1.60 A; A=1-406.
DR PDB; 2ZBZ; X-ray; 1.90 A; A=1-406.
DR PDB; 3CV8; X-ray; 2.00 A; A=1-406.
DR PDB; 3CV9; X-ray; 1.70 A; A=1-406.
DR PDB; 5X7E; X-ray; 1.90 A; A=1-406.
DR PDBsum; 2ZBX; -.
DR PDBsum; 2ZBY; -.
DR PDBsum; 2ZBZ; -.
DR PDBsum; 3CV8; -.
DR PDBsum; 3CV9; -.
DR PDBsum; 5X7E; -.
DR AlphaFoldDB; P18326; -.
DR SMR; P18326; -.
DR KEGG; ag:AAA26823; -.
DR BRENDA; 1.14.15.22; 6031.
DR EvolutionaryTrace; P18326; -.
DR GO; GO:0005737; C:cytoplasm; IDA:UniProtKB.
DR GO; GO:0020037; F:heme binding; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0016709; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen; IDA:UniProtKB.
DR GO; GO:0070640; P:vitamin D3 metabolic process; IDA:UniProtKB.
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; Direct protein sequencing; Heme; Iron;
KW Metal-binding; Monooxygenase; Oxidoreductase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:15207715,
FT ECO:0000269|PubMed:2345149"
FT CHAIN 2..406
FT /note="Vitamin D3 dihydroxylase"
FT /id="PRO_0000052213"
FT REGION 1..24
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1..18
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 81
FT /ligand="calciol"
FT /ligand_id="ChEBI:CHEBI:28940"
FT /evidence="ECO:0000269|PubMed:18937506"
FT BINDING 103
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:18314962"
FT BINDING 107
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:18314962"
FT BINDING 193
FT /ligand="calciol"
FT /ligand_id="ChEBI:CHEBI:28940"
FT /evidence="ECO:0000269|PubMed:18314962,
FT ECO:0000269|PubMed:18937506"
FT BINDING 236
FT /ligand="calciol"
FT /ligand_id="ChEBI:CHEBI:28940"
FT /evidence="ECO:0000269|PubMed:18314962"
FT BINDING 293
FT /ligand="calciol"
FT /ligand_id="ChEBI:CHEBI:28940"
FT /evidence="ECO:0000269|PubMed:18314962,
FT ECO:0000269|PubMed:18937506"
FT BINDING 297
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:18314962"
FT BINDING 353
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /evidence="ECO:0000269|PubMed:18314962"
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:0000269|PubMed:18314962"
FT MUTAGEN 73
FT /note="R->A,F,L,V: Increase of the hydroxylase activity and
FT decrease of affinity for both 25-hydroxyvitamin D3 and 1-
FT alpha-hydroxyvitamin D3. Increase of the hydroxylase
FT activity; when associated with A-84 and F-84."
FT /evidence="ECO:0000269|PubMed:18314962,
FT ECO:0000269|PubMed:18937506"
FT MUTAGEN 84
FT /note="R->A,Q,L,F: Increase of the hydroxylase activity and
FT decrease of affinity for both 25-hydroxyvitamin D3 and 1-
FT alpha-hydroxyvitamin D3. Increase of the hydroxylase
FT activity; when associated with A-73 and V-73."
FT /evidence="ECO:0000269|PubMed:18314962"
FT MUTAGEN 84
FT /note="R->F: Alters the substrate specificity that gives
FT preference to the 1-alpha-hydroxylation of 25-
FT hydroxyvitamin D3 over the 25-hydroxylation of 1-alpha-
FT hydroxyvitamin D3. Increase of the hydroxylase activity and
FT decrease of affinity for both 25-hydroxyvitamin D3 and 1-
FT alpha-hydroxyvitamin D3."
FT /evidence="ECO:0000269|PubMed:18937506"
FT MUTAGEN 88
FT /note="V->A: Decrease of the hydroxylase activity for both
FT 25-hydroxyivitamin D3 and 1-alpha-hydroxyvitamin D3."
FT /evidence="ECO:0000269|PubMed:18314962"
FT MUTAGEN 180
FT /note="L->A: Decrease of the hydroxylase activity for both
FT 25-hydroxyvitamin D3 and 1-alpha-hydroxyvitamin D3."
FT /evidence="ECO:0000269|PubMed:18314962"
FT MUTAGEN 181
FT /note="V->A: Decrease of the hydroxylase activity for both
FT 25-hydroxyvitamin D3 and 1-alpha-hydroxyvitamin D3."
FT /evidence="ECO:0000269|PubMed:18314962"
FT MUTAGEN 193
FT /note="R->A,Q,K: Decrease of the hydroxylase activity."
FT /evidence="ECO:0000269|PubMed:18314962"
FT MUTAGEN 293
FT /note="I->A: Slight increase of the hydroxylase activity."
FT /evidence="ECO:0000269|PubMed:18314962"
FT STRAND 15..17
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 28..35
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 36..44
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 50..54
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 57..64
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 83..85
FT /evidence="ECO:0007829|PDB:2ZBX"
FT TURN 88..90
FT /evidence="ECO:0007829|PDB:2ZBY"
FT HELIX 95..97
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 102..107
FT /evidence="ECO:0007829|PDB:2ZBX"
FT TURN 108..110
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 111..114
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 116..140
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 142..145
FT /evidence="ECO:0007829|PDB:2ZBY"
FT HELIX 146..149
FT /evidence="ECO:0007829|PDB:2ZBX"
FT TURN 150..152
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 153..163
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 167..169
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 170..182
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 186..209
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 215..221
FT /evidence="ECO:0007829|PDB:2ZBX"
FT TURN 222..227
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 231..261
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 264..272
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 274..276
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 277..288
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 292..294
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 296..301
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 303..305
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 308..310
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 315..318
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 320..323
FT /evidence="ECO:0007829|PDB:2ZBX"
FT TURN 327..329
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 330..332
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 358..375
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 380..383
FT /evidence="ECO:0007829|PDB:2ZBX"
FT HELIX 385..387
FT /evidence="ECO:0007829|PDB:2ZBX"
FT STRAND 398..400
FT /evidence="ECO:0007829|PDB:3CV9"
FT STRAND 403..405
FT /evidence="ECO:0007829|PDB:2ZBX"
SQ SEQUENCE 406 AA; 44212 MW; B41CDD7B9BC56191 CRC64;
MTDTATTPQT TDAPAFPSNR SCPYQLPDGY AQLRDTPGPL HRVTLYDGRQ AWVVTKHEAA
RKLLGDPRLS SNRTDDNFPA TSPRFEAVRE SPQAFIGLDP PEHGTRRRMT ISEFTVKRIK
GMRPEVEEVV HGFLDEMLAA GPTADLVSQF ALPVPSMVIC RLLGVPYADH EFFQDASKRL
VQSTDAQSAL TARNDLAGYL DGLITQFQTE PGAGLVGALV ADQLANGEID REELISTAML
LLIAGHETTA SMTSLSVITL LDHPEQYAAL RADRSLVPGA VEELLRYLAI ADIAGGRVAT
ADIEVEGHLI RAGEGVIVVN SIANRDGTVY EDPDALDIHR SARHHLAFGF GVHQCLGQNL
ARLELEVILN ALMDRVPTLR LAVPVEQLVL RPGTTIQGVN ELPVTW
