DNRF_STREF
ID DNRF_STREF Reviewed; 535 AA.
AC Q54530;
DT 19-MAR-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 97.
DE RecName: Full=Aklavinone 12-hydroxylase RdmE;
DE EC=1.14.13.180;
DE AltName: Full=Aklavinone 11-hydroxylase;
GN Name=rdmE;
OS Streptomyces purpurascens.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1924;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 25489 / DSM 40310 / JCM 4509 / NBRC 13077 / Maria 515;
RX PubMed=8081500; DOI=10.1099/00221287-140-6-1351;
RA Niemi J., Ylihonko K., Hakala J., Parssinen R., Kopio A., Mantsala P.;
RT "Hybrid anthracycline antibiotics: production of new anthracyclines by
RT cloned genes from Streptomyces purpurascens in Streptomyces galilaeus.";
RL Microbiology 140:1351-1358(1994).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 25489 / DSM 40310 / JCM 4509 / NBRC 13077 / Maria 515;
RA Halo L., Wang Y., Mantsala P., Hakala J., Ylihonko K.;
RT "Characterization of the gene cluster involved in rhodomycin
RT biosynthesis.";
RL Submitted (AUG-1994) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 25489 / DSM 40310 / JCM 4509 / NBRC 13077 / Maria 515;
RX PubMed=7751313; DOI=10.1128/jb.177.10.2942-2945.1995;
RA Niemi J., Mantsala P.;
RT "Nucleotide sequences and expression of genes from Streptomyces
RT purpurascens that cause the production of new anthracyclines in
RT Streptomyces galilaeus.";
RL J. Bacteriol. 177:2942-2945(1995).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 25489 / DSM 40310 / JCM 4509 / NBRC 13077 / Maria 515;
RA Halo L., Wang Y., Mantsala P., Hakala J., Ylihonko K.;
RL Submitted (OCT-2001) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP PROTEIN SEQUENCE OF 2-13, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION, AND SUBUNIT.
RX PubMed=10082933; DOI=10.1016/s0167-4838(98)00265-9;
RA Niemi J., Wang Y., Airas K., Ylihonko K., Hakala J., Mantsala P.;
RT "Characterization of aklavinone-11-hydroxylase from Streptomyces
RT purpurascens.";
RL Biochim. Biophys. Acta 1430:57-64(1999).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.49 ANGSTROMS) IN COMPLEX WITH FAD AND AKLAVINONE,
RP FUNCTION, ACTIVE SITE, MUTAGENESIS OF TYR-224 AND ARG-373, SUBSTRATE
RP SPECIFICITY, REACTION MECHANISM, COFACTOR, AND SUBUNIT.
RX PubMed=19744497; DOI=10.1016/j.jmb.2009.09.003;
RA Lindqvist Y., Koskiniemi H., Jansson A., Sandalova T., Schnell R., Liu Z.,
RA Mantsala P., Niemi J., Schneider G.;
RT "Structural basis for substrate recognition and specificity in aklavinone-
RT 11-hydroxylase from rhodomycin biosynthesis.";
RL J. Mol. Biol. 393:966-977(2009).
CC -!- FUNCTION: Involved in the biosynthesis of the anthracyclines
CC carminomycin, rhodomycin and daunorubicin (daunomycin) which are
CC aromatic polyketide antibiotics that exhibit high cytotoxicity and are
CC widely applied in the chemotherapy of a variety of cancers. Catalyzes
CC the incorporation of a hydroxyl group at position C-11 of aklavinone,
CC resulting in epsilon-rhodomycinone. It cannot accept substrates
CC glycosylated at position C-7 and is specific for the C-9R configuration
CC of anthracyclines. It can use both NAD or NADP but it is slowly
CC inactivated in the presence of NADH. {ECO:0000269|PubMed:10082933,
CC ECO:0000269|PubMed:19744497}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=aklavinone + H(+) + NADPH + O2 = epsilon-rhodomycinone + H2O +
CC NADP(+); Xref=Rhea:RHEA:37835, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:31181, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:75291; EC=1.14.13.180;
CC Evidence={ECO:0000269|PubMed:10082933};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:19744497};
CC -!- ACTIVITY REGULATION: Inhibited by phenylglyoxal and 2,3-butanedione.
CC NADP provides a partial protection against inhibition by phenylglyoxal.
CC Increasing the methanol concentration in the assay causes inhibition of
CC the enzyme. {ECO:0000269|PubMed:10082933}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2 uM for NAD (at pH 7.5) {ECO:0000269|PubMed:10082933};
CC KM=10 uM for aklavinone (at pH 7.5) {ECO:0000269|PubMed:10082933};
CC KM=2 mM for NADP (at pH 7.5) {ECO:0000269|PubMed:10082933};
CC pH dependence:
CC Optimum pH is near 7.4. {ECO:0000269|PubMed:10082933};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius.
CC {ECO:0000269|PubMed:10082933};
CC -!- PATHWAY: Antibiotic biosynthesis; daunorubicin biosynthesis.
CC -!- PATHWAY: Antibiotic biosynthesis; carminomycin biosynthesis.
CC -!- PATHWAY: Antibiotic biosynthesis; rhodomycin biosynthesis.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:10082933,
CC ECO:0000269|PubMed:19744497}.
CC -!- SIMILARITY: Belongs to the PheA/TfdB FAD monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; U10405; AAA83424.1; -; Genomic_DNA.
DR PDB; 3IHG; X-ray; 2.49 A; A/B/C=1-535.
DR PDBsum; 3IHG; -.
DR AlphaFoldDB; Q54530; -.
DR SMR; Q54530; -.
DR KEGG; ag:AAA83424; -.
DR BRENDA; 1.14.13.180; 6079.
DR UniPathway; UPA00054; -.
DR UniPathway; UPA01040; -.
DR UniPathway; UPA01042; -.
DR EvolutionaryTrace; Q54530; -.
DR GO; GO:0071949; F:FAD binding; IDA:UniProtKB.
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:1901771; P:daunorubicin biosynthetic process; IDA:UniProtKB.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR002938; FAD-bd.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR Pfam; PF01494; FAD_binding_3; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Direct protein sequencing; FAD;
KW Flavoprotein; NAD; NADP; Nucleotide-binding; Oxidoreductase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:10082933"
FT CHAIN 2..535
FT /note="Aklavinone 12-hydroxylase RdmE"
FT /id="PRO_0000425675"
FT ACT_SITE 224
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:19744497"
FT BINDING 15..16
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:19744497"
FT BINDING 35
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:19744497"
FT BINDING 119
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:19744497"
FT BINDING 143
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:19744497"
FT BINDING 308
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:19744497"
FT BINDING 317
FT /ligand="aklavinone"
FT /ligand_id="ChEBI:CHEBI:31181"
FT /evidence="ECO:0000269|PubMed:19744497"
FT MUTAGEN 224
FT /note="Y->F: Loss of hydroxylase activity."
FT /evidence="ECO:0000269|PubMed:19744497"
FT MUTAGEN 373
FT /note="R->A: Hydroxylase activity similar as the wild-type
FT enzyme."
FT /evidence="ECO:0000269|PubMed:19744497"
FT MUTAGEN 373
FT /note="R->M: Hydroxylase activity similar as the wild-type
FT enzyme."
FT /evidence="ECO:0000269|PubMed:19744497"
FT MUTAGEN 373
FT /note="R->Q: Hydroxylase activity similar as the wild-type
FT enzyme."
FT /evidence="ECO:0000269|PubMed:19744497"
FT STRAND 5..11
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 15..25
FT /evidence="ECO:0007829|PDB:3IHG"
FT TURN 26..28
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 31..34
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 36..39
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 51..59
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 63..68
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 80..89
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 91..97
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 99..104
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 107..109
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 119..132
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 136..140
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 142..149
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 151..153
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 155..163
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 166..177
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 184..188
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 193..208
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 211..213
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 221..226
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 231..236
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 242..249
FT /evidence="ECO:0007829|PDB:3IHG"
FT TURN 251..254
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 257..259
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 262..273
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 281..296
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 298..300
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 303..305
FT /evidence="ECO:0007829|PDB:3IHG"
FT TURN 307..310
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 320..339
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 347..373
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 376..378
FT /evidence="ECO:0007829|PDB:3IHG"
FT TURN 379..381
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 388..392
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 430..433
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 436..439
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 440..443
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 445..452
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 457..470
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 474..479
FT /evidence="ECO:0007829|PDB:3IHG"
FT TURN 480..482
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 490..493
FT /evidence="ECO:0007829|PDB:3IHG"
FT TURN 497..499
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 501..504
FT /evidence="ECO:0007829|PDB:3IHG"
FT STRAND 508..516
FT /evidence="ECO:0007829|PDB:3IHG"
FT HELIX 521..532
FT /evidence="ECO:0007829|PDB:3IHG"
SQ SEQUENCE 535 AA; 57437 MW; 969CAAB6FEB6AD4F CRC64;
MNDHEVDVLV VGAGLGGLST AMFLARQGVR VLVVERRPGL SPYPRAAGQN PRTMELLRIG
GVADEVVRAD DIRGTQGDFV IRLAESVRGE ILRTVSESFD DMVAATEPCT PAGWAMLSQD
KLEPILLAQA RKHGGAIRFG TRLLSFRQHD DDAGAGVTAR LAGPDGEYDL RAGYLVGADG
NRSLVRESLG IGRYGHGTLT HMVGVIFDAD LSGIMEPGTT GWYYLHHPEF KGTFGPTDRP
DRHTLFVEYD PDEGERPEDF TPQRCVELIG LALDAPEVKP ELVDIQGWEM AARIAERWRE
GRVFLAGDAA KVTPPTGGMS GNAAVADGFD LAWKLAAVLQ GQAGAGLLDT YEDERKVAAE
LVVAEALAIY AQRMAPHMAE VWDKSVGYPE TLLGFRYRSS AVLATDDDPA RVENPLTPSG
RPGFRGPHVL VSRHGERLST VDLFGDGWTL LAGELGADWV AAAEAVSAEL GVPVRAYRVG
AGLTDPESAV SERYGIGKAG ASLVRPDGIV AWRTDEAAAD AAQTLEGVLR RVLDR
