CP51A_RAT
ID CP51A_RAT Reviewed; 503 AA.
AC Q64654; Q64549;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 168.
DE RecName: Full=Lanosterol 14-alpha demethylase {ECO:0000303|PubMed:8024575};
DE Short=LDM;
DE EC=1.14.14.154 {ECO:0000269|PubMed:10544287, ECO:0000269|PubMed:11328599, ECO:0000269|PubMed:7665087};
DE AltName: Full=CYPLI;
DE AltName: Full=Cytochrome P450 51A1;
DE AltName: Full=Cytochrome P450-14DM;
DE Short=Cytochrome P45014DM;
DE AltName: Full=Cytochrome P450LI;
DE AltName: Full=Sterol 14-alpha demethylase;
GN Name=Cyp51a1 {ECO:0000303|PubMed:11328599, ECO:0000312|RGD:2481};
GN Synonyms=Cyp51;
OS Rattus norvegicus (Rat).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC Murinae; Rattus.
OX NCBI_TaxID=10116;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RC TISSUE=Liver;
RX PubMed=8024575; DOI=10.1006/bbrc.1994.1848;
RA Aoyama Y., Funae Y., Noshiro M., Horiuchi T., Yoshida Y.;
RT "Occurrence of a P450 showing high homology to yeast lanosterol 14-
RT demethylase (P450(14DM)) in the rat liver.";
RL Biochem. Biophys. Res. Commun. 201:1320-1326(1994).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=Wistar;
RX PubMed=9498553; DOI=10.1093/oxfordjournals.jbchem.a021869;
RA Noshiro M., Aoyama Y., Kawamoto T., Gotoh O., Horiuchi T., Yoshida Y.;
RT "Structural and evolutionary studies on sterol 14-demethylase P450 (CYP51),
RT the most conserved P450 monooxygenase: I. Structural analyses of the gene
RT and multiple sizes of mRNA.";
RL J. Biochem. 122:1114-1121(1997).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC TISSUE=Kidney;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA project:
RT the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [4]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 60-78; 223-231; 271-285;
RP 386-405; 437-446 AND 450-461, FUNCTION, CATALYTIC ACTIVITY, AND ACTIVITY
RP REGULATION.
RC TISSUE=Liver;
RX PubMed=7665087; DOI=10.1016/0378-1119(95)00211-n;
RA Sloane D.L., So O.Y., Leung R., Scarafia L.E., Saldou N., Jarnagin K.,
RA Swinney D.C.;
RT "Cloning and functional expression of the cDNA encoding rat lanosterol 14-
RT alpha demethylase.";
RL Gene 161:243-248(1995).
RN [5]
RP PROTEIN SEQUENCE OF 52-81; 305-319; 324-334; 467-482 AND 488-499, AND
RP SUBCELLULAR LOCATION.
RC TISSUE=Liver;
RX PubMed=8797093; DOI=10.1093/oxfordjournals.jbchem.a021331;
RA Aoyama Y., Noshiro M., Gotoh O., Imaoka S., Funae Y., Kurosawa N.,
RA Horiuchi T., Yoshida Y.;
RT "Sterol 14-demethylase P450 (P45014DM*) is one of the most ancient and
RT conserved P450 species.";
RL J. Biochem. 119:926-933(1996).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY
RP REGULATION.
RX PubMed=10544287; DOI=10.1093/oxfordjournals.jbchem.a022536;
RA Nitahara Y., Aoyama Y., Horiuchi T., Noshiro M., Yoshida Y.;
RT "Purification and characterization of rat sterol 14-demethylase P450
RT (CYP51) expressed in Escherichia coli.";
RL J. Biochem. 126:927-933(1999).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF VAL-130; THR-136; PHE-139;
RP ALA-144; TYR-227; ASP-231; HIS-314; THR-315 AND SER-316.
RX PubMed=11328599; DOI=10.1093/oxfordjournals.jbchem.a002917;
RA Nitahara Y., Kishimoto K., Yabusaki Y., Gotoh O., Yoshida Y., Horiuchi T.,
RA Aoyama Y.;
RT "The amino acid residues affecting the activity and azole susceptibility of
RT rat CYP51 (sterol 14-demethylase P450).";
RL J. Biochem. 129:761-768(2001).
CC -!- FUNCTION: A cytochrome P450 monooxygenase involved in sterol
CC biosynthesis. Catalyzes 14-alpha demethylation of lanosterol and 24,25-
CC dihydrolanosterol likely through sequential oxidative conversion of 14-
CC alpha methyl group to hydroxymethyl, then to carboxylaldehyde, followed
CC by the formation of the delta 14,15 double bond in the sterol core and
CC concomitant release of formic acid (PubMed:7665087, PubMed:10544287,
CC PubMed:11328599). Mechanistically, uses molecular oxygen inserting one
CC oxygen atom into a substrate, and reducing the second into a water
CC molecule, with two electrons provided by NADPH via cytochrome P450
CC reductase (CPR; NADPH-ferrihemoprotein reductase) (PubMed:7665087,
CC PubMed:10544287, PubMed:11328599). {ECO:0000269|PubMed:10544287,
CC ECO:0000269|PubMed:11328599, ECO:0000269|PubMed:7665087}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 14alpha-methyl steroid + 3 O2 + 3 reduced [NADPH--
CC hemoprotein reductase] = a Delta(14) steroid + formate + 4 H(+) + 4
CC H2O + 3 oxidized [NADPH--hemoprotein reductase];
CC Xref=Rhea:RHEA:54028, Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:15740, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC ChEBI:CHEBI:138029, ChEBI:CHEBI:138031; EC=1.14.14.154;
CC Evidence={ECO:0000269|PubMed:10544287, ECO:0000269|PubMed:11328599,
CC ECO:0000269|PubMed:7665087};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54029;
CC Evidence={ECO:0000305|PubMed:10544287, ECO:0000305|PubMed:11328599};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=lanosterol + 3 O2 + 3 reduced [NADPH--hemoprotein reductase] =
CC 4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol + formate + 4
CC H(+) + 4 H2O + 3 oxidized [NADPH--hemoprotein reductase];
CC Xref=Rhea:RHEA:25286, Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:15740, ChEBI:CHEBI:16521, ChEBI:CHEBI:17813,
CC ChEBI:CHEBI:57618, ChEBI:CHEBI:58210; EC=1.14.14.154;
CC Evidence={ECO:0000269|PubMed:10544287, ECO:0000269|PubMed:11328599};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25287;
CC Evidence={ECO:0000305|PubMed:10544287, ECO:0000305|PubMed:11328599};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=24,25-dihydrolanosterol + 3 O2 + 3 reduced [NADPH--hemoprotein
CC reductase] = 4,4-dimethyl-8,14-cholestadien-3beta-ol + formate + 4
CC H(+) + 4 H2O + 3 oxidized [NADPH--hemoprotein reductase];
CC Xref=Rhea:RHEA:45960, Rhea:RHEA-COMP:11964, Rhea:RHEA-COMP:11965,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:15740, ChEBI:CHEBI:28113, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210, ChEBI:CHEBI:78904;
CC Evidence={ECO:0000269|PubMed:10544287, ECO:0000269|PubMed:7665087};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:45961;
CC Evidence={ECO:0000305|PubMed:10544287};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q16850};
CC -!- ACTIVITY REGULATION: Inhibited by azalanstat (PubMed:7665087).
CC Inhibited by azole antifungal agents ketoconazole, itraconazole and
CC fluconazole (PubMed:10544287). {ECO:0000269|PubMed:10544287,
CC ECO:0000269|PubMed:7665087}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=10.5 uM for lanosterol {ECO:0000269|PubMed:10544287};
CC KM=20.0 uM for 24,25-dihydrolanosterol {ECO:0000269|PubMed:10544287};
CC Vmax=13.9 nmol/min/nmol enzyme toward lanosterol
CC {ECO:0000269|PubMed:10544287};
CC Vmax=20.0 nmol/min/nmol enzyme toward 24,25-dihydrolanosterol
CC {ECO:0000269|PubMed:10544287};
CC -!- PATHWAY: Steroid biosynthesis; zymosterol biosynthesis; zymosterol from
CC lanosterol: step 1/6. {ECO:0000305|PubMed:7665087}.
CC -!- SUBCELLULAR LOCATION: Endoplasmic reticulum membrane
CC {ECO:0000305|PubMed:8797093}; Single-pass membrane protein
CC {ECO:0000255}. Microsome membrane {ECO:0000269|PubMed:8797093}; Single-
CC pass membrane protein {ECO:0000255}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAA87074.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; D55681; BAA09529.1; -; mRNA.
DR EMBL; AB004096; BAA20354.1; -; Genomic_DNA.
DR EMBL; BC087033; AAH87033.1; -; mRNA.
DR EMBL; U17697; AAA87074.1; ALT_FRAME; mRNA.
DR PIR; JC4240; JC4240.
DR PIR; JC4758; JC4758.
DR RefSeq; NP_037073.1; NM_012941.2.
DR AlphaFoldDB; Q64654; -.
DR SMR; Q64654; -.
DR BioGRID; 247462; 1.
DR STRING; 10116.ENSRNOP00000009985; -.
DR BindingDB; Q64654; -.
DR ChEMBL; CHEMBL4981; -.
DR DrugCentral; Q64654; -.
DR SwissLipids; SLP:000001302; -.
DR jPOST; Q64654; -.
DR PaxDb; Q64654; -.
DR PRIDE; Q64654; -.
DR Ensembl; ENSRNOT00000009985; ENSRNOP00000009985; ENSRNOG00000007234.
DR GeneID; 25427; -.
DR KEGG; rno:25427; -.
DR UCSC; RGD:2481; rat.
DR CTD; 13121; -.
DR RGD; 2481; Cyp51.
DR eggNOG; KOG0684; Eukaryota.
DR GeneTree; ENSGT00930000151026; -.
DR HOGENOM; CLU_001570_15_0_1; -.
DR InParanoid; Q64654; -.
DR OMA; AWTLIEL; -.
DR OrthoDB; 572303at2759; -.
DR PhylomeDB; Q64654; -.
DR TreeFam; TF105091; -.
DR BRENDA; 1.14.14.154; 5301.
DR Reactome; R-RNO-191273; Cholesterol biosynthesis.
DR Reactome; R-RNO-211976; Endogenous sterols.
DR UniPathway; UPA00770; UER00754.
DR PRO; PR:Q64654; -.
DR Proteomes; UP000002494; Chromosome 4.
DR Bgee; ENSRNOG00000007234; Expressed in duodenum and 21 other tissues.
DR Genevisible; Q64654; RN.
DR GO; GO:0001669; C:acrosomal vesicle; IDA:RGD.
DR GO; GO:0005737; C:cytoplasm; IDA:RGD.
DR GO; GO:0005789; C:endoplasmic reticulum membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0043231; C:intracellular membrane-bounded organelle; IDA:RGD.
DR GO; GO:0020037; F:heme binding; ISO:RGD.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IBA:GO_Central.
DR GO; GO:0016491; F:oxidoreductase activity; IBA:GO_Central.
DR GO; GO:0008398; F:sterol 14-demethylase activity; IDA:RGD.
DR GO; GO:0033488; P:cholesterol biosynthetic process via 24,25-dihydrolanosterol; IDA:RGD.
DR GO; GO:1900222; P:negative regulation of amyloid-beta clearance; ISO:RGD.
DR GO; GO:0042177; P:negative regulation of protein catabolic process; ISO:RGD.
DR GO; GO:0050709; P:negative regulation of protein secretion; ISO:RGD.
DR GO; GO:0060282; P:positive regulation of oocyte development; IMP:RGD.
DR GO; GO:0045540; P:regulation of cholesterol biosynthetic process; TAS:RGD.
DR GO; GO:0044752; P:response to human chorionic gonadotropin; IEP:RGD.
DR GO; GO:0032868; P:response to insulin; IEP:RGD.
DR GO; GO:0010288; P:response to lead ion; IEP:RGD.
DR GO; GO:0007283; P:spermatogenesis; IEP:RGD.
DR GO; GO:0006694; P:steroid biosynthetic process; ISO:RGD.
DR GO; GO:0016125; P:sterol metabolic process; IBA:GO_Central.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002403; Cyt_P450_E_grp-IV.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00465; EP450IV.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Cholesterol biosynthesis; Cholesterol metabolism;
KW Direct protein sequencing; Endoplasmic reticulum; Heme; Iron;
KW Lipid biosynthesis; Lipid metabolism; Membrane; Metal-binding; Microsome;
KW Monooxygenase; Oxidoreductase; Reference proteome; Steroid biosynthesis;
KW Steroid metabolism; Sterol biosynthesis; Sterol metabolism; Transmembrane;
KW Transmembrane helix.
FT CHAIN 1..503
FT /note="Lanosterol 14-alpha demethylase"
FT /id="PRO_0000052001"
FT TRANSMEM 24..44
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 449
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|UniProtKB:Q16850"
FT MUTAGEN 130
FT /note="V->A: Decreases lanosterol 14-alpha demethylase
FT activity."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 136
FT /note="T->A,V: Decreases lanosterol 14-alpha demethylase
FT activity."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 139
FT /note="F->A: Impairs lanosterol 14-alpha demethylase
FT activity."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 144
FT /note="A->I,V: Decreases lanosterol 14-alpha demethylase
FT activity. Increases the susceptibility to ketoconazole."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 227
FT /note="Y->A: Significatly decreases lanosterol 14-alpha
FT demethylase activity. Increases the susceptibility to
FT ketoconazole."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 231
FT /note="D->A,E: Significatly decreases lanosterol 14-alpha
FT demethylase activity."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 314
FT /note="H->F,A,K,D: Significatly decreases lanosterol 14-
FT alpha demethylase activity. Increases the susceptibility to
FT ketoconazole."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 315
FT /note="T->A,V,K,N: Impairs lanosterol 14-alpha demethylase
FT activity. Increases the susceptibility to ketoconazole."
FT /evidence="ECO:0000269|PubMed:11328599"
FT MUTAGEN 316
FT /note="S->T,V,L: Significatly decreases lanosterol 14-alpha
FT demethylase activity."
FT /evidence="ECO:0000269|PubMed:11328599"
FT CONFLICT 181
FT /note="E -> K (in Ref. 4; AAA87074)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 503 AA; 56707 MW; 33D8F345FFE9CF21 CRC64;
MVLLGLLQSG GSVLGQAMEQ VTGGNLLSTL LIACAFTLSL VYLFRLAVGH MVQLPAGAKS
PPYIYSPIPF LGHAIAFGKS PIEFLENAYE KYGPVFSFTM VGKTFTYLLG SDAAALLFNS
KNEDLNAEEV YGRLTTPVFG KGVAYDVPNA VFLEQKKILK SGLNIAHFKQ YVSIIEKEAK
EYFKSWGESG ERNVFEALSE LIILTASHCL HGKEIRSQLN EKVAQLYADL DGGFSHAAWL
LPGWLPLPSF RRRDRAHREI KNIFYKAIQK RRLSKEPAED ILQTLLDSTY KDGRPLTDDE
IAGMLIGLLL AGQHTSSTTS AWMGFFLARD KPLQDKCYLE QKTVCGEDLP PLTYEQLKDL
NLLDRCIKET LRLRPPIMTM MRMAKTPQTV AGYTIPPGHQ VCVSPTVNQR LKDSWVERLD
FNPDRYLQDN PASGEKFAYV PFGAGRHRCI GENFAYVQIK TIWSTMLRLY EFDLINGYFP
SVNYTTMIHT PENPVIRYKR RSK
