5BPOR_DIGLA
ID 5BPOR_DIGLA Reviewed; 389 AA.
AC Q6PQJ9;
DT 28-NOV-2012, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 70.
DE RecName: Full=3-oxo-Delta(4,5)-steroid 5-beta-reductase;
DE EC=1.3.1.3 {ECO:0000269|PubMed:16386278, ECO:0000269|PubMed:18032383};
DE AltName: Full=Delta(4)-3-oxosteroid 5-beta-reductase;
DE AltName: Full=Delta-4,5-steroid 5-beta-reductase;
DE Short=At5beta-StR;
DE AltName: Full=Progesterone 5-beta-reductase;
DE Short=5beta-POR;
OS Digitalis lanata (Grecian foxglove).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Lamiales; Plantaginaceae; Digitalideae; Digitalis.
OX NCBI_TaxID=49450;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP SUBSTRATE SPECIFICITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC TISSUE=Leaf;
RX PubMed=16386278; DOI=10.1016/j.phytochem.2005.11.013;
RA Herl V., Fischer G., Muller-Uri F., Kreis W.;
RT "Molecular cloning and heterologous expression of progesterone 5beta-
RT reductase from Digitalis lanata Ehrh.";
RL Phytochemistry 67:225-231(2006).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC TISSUE=Leaf;
RX AGRICOLA=IND44043462; DOI=10.1007/s00606-007-0616-0;
RA Herl V., Albach D.C., Mueller-Uri F., Braeuchler C., Heubl G., Kreis W.;
RT "Using progesterone 5beta-reductase, a gene encoding a key enzyme in the
RT cardenolide biosynthesis, to infer the phylogeny of the genus Digitalis.";
RL Plant Syst. Evol. 271:65-78(2008).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 26-389 IN COMPLEX WITH NADP,
RP SUBUNIT, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITES, AND MUTAGENESIS OF
RP TYR-179.
RX PubMed=18032383; DOI=10.1074/jbc.m706185200;
RA Thorn A., Egerer-Sieber C., Jager C.M., Herl V., Muller-Uri F., Kreis W.,
RA Muller Y.A.;
RT "The crystal structure of progesterone 5beta-reductase from Digitalis
RT lanata defines a novel class of short chain dehydrogenases/reductases.";
RL J. Biol. Chem. 283:17260-17269(2008).
CC -!- FUNCTION: Involved in cardenolide biosynthesis. Catalyzes the
CC stereospecific conversion of progesterone to 5-beta-pregnane-3,20-
CC dione. Can use progesterone, testosterone, 4-androstene-3,17-dione,
CC cortisol and cortisone as substrates, but not pregnenolone, 21-OH-
CC pregnenolone or isoprogesterone. NADPH could not be replaced by NADH.
CC {ECO:0000269|PubMed:16386278, ECO:0000269|PubMed:18032383}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5beta-cholestan-3-one + NADP(+) = cholest-4-en-3-one + H(+) +
CC NADPH; Xref=Rhea:RHEA:11524, ChEBI:CHEBI:15378, ChEBI:CHEBI:16074,
CC ChEBI:CHEBI:16175, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.3.1.3;
CC Evidence={ECO:0000269|PubMed:16386278, ECO:0000269|PubMed:18032383};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4,5beta-dihydrocortisone + NADP(+) = cortisone + H(+) + NADPH;
CC Xref=Rhea:RHEA:14037, ChEBI:CHEBI:15378, ChEBI:CHEBI:16962,
CC ChEBI:CHEBI:18093, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.3.1.3;
CC Evidence={ECO:0000269|PubMed:16386278, ECO:0000269|PubMed:18032383};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=120 uM for progesterone {ECO:0000269|PubMed:16386278};
CC KM=291 uM for cortisol {ECO:0000269|PubMed:16386278};
CC KM=228 uM for 4-androstene-3,17-dione {ECO:0000269|PubMed:16386278};
CC KM=1.597 mM for cortexone {ECO:0000269|PubMed:16386278};
CC KM=8 uM for NADPH {ECO:0000269|PubMed:16386278};
CC Vmax=45 nmol/sec/mg enzyme with progesterone as substrate
CC {ECO:0000269|PubMed:16386278};
CC Vmax=81.2 nmol/sec/mg enzyme with cortisol as substrate
CC {ECO:0000269|PubMed:16386278};
CC Vmax=18.5 nmol/sec/mg enzyme with 4-androstene-3,17-dione as
CC substrate {ECO:0000269|PubMed:16386278};
CC Vmax=63.3 nmol/sec/mg enzyme with cortexone as substrate
CC {ECO:0000269|PubMed:16386278};
CC Vmax=31.1 nmol/sec/mg enzyme with NADPH as substrate
CC {ECO:0000269|PubMed:16386278};
CC pH dependence:
CC Optimum pH is 7.8. {ECO:0000269|PubMed:16386278};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:16386278};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:18032383}.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. Highly divergent. {ECO:0000305}.
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DR EMBL; AY574950; AAS76634.1; -; mRNA.
DR EMBL; AY585867; AAS93804.1; -; Genomic_DNA.
DR PDB; 2V6F; X-ray; 2.40 A; A=26-389.
DR PDB; 2V6G; X-ray; 2.30 A; A=26-389.
DR PDBsum; 2V6F; -.
DR PDBsum; 2V6G; -.
DR AlphaFoldDB; Q6PQJ9; -.
DR SMR; Q6PQJ9; -.
DR BioCyc; MetaCyc:MON-14261; -.
DR BRENDA; 1.3.1.3; 1947.
DR EvolutionaryTrace; Q6PQJ9; -.
DR GO; GO:0047787; F:delta4-3-oxosteroid 5beta-reductase activity; IEA:UniProtKB-EC.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR InterPro; IPR001509; Epimerase_deHydtase.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF01370; Epimerase; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; NADP; Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..389
FT /note="3-oxo-Delta(4,5)-steroid 5-beta-reductase"
FT /id="PRO_0000420239"
FT ACT_SITE 147
FT /evidence="ECO:0000269|PubMed:18032383"
FT ACT_SITE 179
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 35..37
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 63..64
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 81..82
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 105
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 143
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 179
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 206
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT BINDING 213..215
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:18032383"
FT MUTAGEN 179
FT /note="Y->A,F: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:18032383"
FT STRAND 27..33
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 37..45
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 55..64
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 76..79
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 85..92
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 100..103
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 112..130
FT /evidence="ECO:0007829|PDB:2V6G"
FT TURN 131..133
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 139..143
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 147..150
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 153..155
FT /evidence="ECO:0007829|PDB:2V6G"
FT TURN 156..158
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 164..166
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 168..170
FT /evidence="ECO:0007829|PDB:2V6F"
FT HELIX 178..190
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 197..207
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 217..231
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 241..245
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 253..265
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 267..269
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 272..276
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 284..295
FT /evidence="ECO:0007829|PDB:2V6G"
FT STRAND 304..306
FT /evidence="ECO:0007829|PDB:2V6F"
FT HELIX 310..313
FT /evidence="ECO:0007829|PDB:2V6G"
FT TURN 314..316
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 318..327
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 335..338
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 341..348
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 358..362
FT /evidence="ECO:0007829|PDB:2V6G"
FT HELIX 371..384
FT /evidence="ECO:0007829|PDB:2V6G"
SQ SEQUENCE 389 AA; 44071 MW; 184B9EDCDDC26E04 CRC64;
MSWWWAGAIG AAKKRLEEDD AQPKHSSVAL IVGVTGIIGN SLAEILPLAD TPGGPWKVYG
VARRTRPAWH EDNPINYVQC DISDPDDSQA KLSPLTDVTH VFYVTWANRS TEQENCEANS
KMFRNVLDAV IPNCPNLKHI SLQTGRKHYM GPFESYGKIE SHDPPYTEDL PRLKYMNFYY
DLEDIMLEEV EKKEGLTWSV HRPGNIFGFS PYSMMNLVGT LCVYAAICKH EGKVLRFTGC
KAAWDGYSDC SDADLIAEHH IWAAVDPYAK NEAFNVSNGD VFKWKHFWKV LAEQFGVGCG
EYEEGVDLKL QDLMKGKEPV WEEIVRENGL TPTKLKDVGI WWFGDVILGN ECFLDSMNKS
KEHGFLGFRN SKNAFISWID KAKAYKIVP
