(data stored in SCRATCH zone)

SWISSPROT: ERYK_SACEN

ID   ERYK_SACEN              Reviewed;         397 AA.
AC   P48635; A4F7N0; O33990;
DT   01-FEB-1996, integrated into UniProtKB/Swiss-Prot.
DT   22-FEB-2012, sequence version 3.
DT   11-DEC-2019, entry version 107.
DE   RecName: Full=Erythromycin C-12 hydroxylase;
DE            EC=1.14.13.154;
DE   AltName: Full=Cytochrome P450 113A1;
DE            Short=CYP113A1;
DE   AltName: Full=Erythromycin D C-12 hydroxylase;
GN   Name=eryK; Synonyms=CYP113A1; OrderedLocusNames=SACE_0713;
OS   Saccharopolyspora erythraea (strain ATCC 11635 / DSM 40517 / JCM 4748 /
OS   NBRC 13426 / NCIMB 8594 / NRRL 2338).
OC   Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae;
OC   Saccharopolyspora.
OX   NCBI_TaxID=405948;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION IN ERYTHROMYCIN BIOSYNTHESIS,
RP   AND DISRUPTION PHENOTYPE.
RC   STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC   2338;
RX   PubMed=8416893; DOI=10.1128/jb.175.1.182-189.1993;
RA   Stassi D., Donadio S., Staver M.J., Katz L.;
RT   "Identification of a Saccharopolyspora erythraea gene required for the
RT   final hydroxylation step in erythromycin biosynthesis.";
RL   J. Bacteriol. 175:182-189(1993).
RN   [2]
RP   SEQUENCE REVISION.
RC   STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC   2338;
RX   PubMed=9249068; DOI=10.1016/s0378-1119(97)00086-3;
RA   Pereda A., Summers R., Katz L.;
RT   "Nucleotide sequence of the ermE distal flank of the erythromycin
RT   biosynthesis cluster in Saccharopolyspora erythraea.";
RL   Gene 193:65-71(1997).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC   2338;
RX   PubMed=17369815; DOI=10.1038/nbt1297;
RA   Oliynyk M., Samborskyy M., Lester J.B., Mironenko T., Scott N., Dickens S.,
RA   Haydock S.F., Leadlay P.F.;
RT   "Complete genome sequence of the erythromycin-producing bacterium
RT   Saccharopolyspora erythraea NRRL23338.";
RL   Nat. Biotechnol. 25:447-453(2007).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE SPECIFICITY, KINETIC
RP   PARAMETERS, PATHWAY, AND SUBUNIT.
RC   STRAIN=ATCC 11635 / DSM 40517 / JCM 4748 / NBRC 13426 / NCIMB 8594 / NRRL
RC   2338;
RX   PubMed=7849045; DOI=10.1021/bi00006a006;
RA   Lambalot R.H., Cane D.E., Aparicio J.J., Katz L.;
RT   "Overproduction and characterization of the erythromycin C-12 hydroxylase,
RT   EryK.";
RL   Biochemistry 34:1858-1866(1995).
RN   [5]
RP   CRYSTALLIZATION.
RX   PubMed=19075827; DOI=10.2174/092986608786071201;
RA   Savino C., Sciara G., Miele A.E., Kendrew S.G., Vallone B.;
RT   "Cloning, expression, purification, crystallization and preliminary X-ray
RT   crystallographic analysis of C-12 hydroxylase EryK from Saccharopolyspora
RT   erythraea.";
RL   Protein Pept. Lett. 15:1138-1141(2008).
RN   [6]
RP   X-RAY CRYSTALLOGRAPHY (1.59 ANGSTROMS) OF 2-397 IN COMPLEXES WITH HEME AND
RP   ERYTHROMYCIN D, COFACTOR, AND SUBUNIT.
RX   PubMed=19625248; DOI=10.1074/jbc.m109.003590;
RA   Savino C., Montemiglio L.C., Sciara G., Miele A.E., Kendrew S.G., Jemth P.,
RA   Gianni S., Vallone B.;
RT   "Investigating the structural plasticity of a cytochrome P450: three-
RT   dimensional structures of P450 EryK and binding to its physiological
RT   substrate.";
RL   J. Biol. Chem. 284:29170-29179(2009).
RN   [7]
RP   X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF 2-397 IN COMPLEXES WITH HEME AND
RP   THE INHIBITORS KETOCONAZOLE AND CLOTRIMAZOLE, AND COFACTOR.
RX   PubMed=20845962; DOI=10.1021/bi101062v;
RA   Montemiglio L.C., Gianni S., Vallone B., Savino C.;
RT   "Azole drugs trap cytochrome P450 EryK in alternative conformational
RT   states.";
RL   Biochemistry 49:9199-9206(2010).
CC   -!- FUNCTION: Responsible for the C-12 hydroxylation of the macrolactone
CC       ring of erythromycin. Thus, EryK catalyzes the hydroxylation of
CC       erythromycin D (ErD) at the C-12 position to produce erythromycin C
CC       (ErC). Erythromycin B (ErB) is not a substrate for this enzyme.
CC       {ECO:0000269|PubMed:7849045, ECO:0000269|PubMed:8416893}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=erythromycin D + H(+) + NADPH + O2 = erythromycin C + H2O +
CC         NADP(+); Xref=Rhea:RHEA:32631, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:15379, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC         ChEBI:CHEBI:63677, ChEBI:CHEBI:64258; EC=1.14.13.154;
CC         Evidence={ECO:0000269|PubMed:7849045};
CC   -!- COFACTOR:
CC       Name=heme b; Xref=ChEBI:CHEBI:60344;
CC         Evidence={ECO:0000269|PubMed:19625248, ECO:0000269|PubMed:20845962,
CC         ECO:0000269|PubMed:7849045};
CC       Note=Binds 1 heme b (iron(II)-protoporphyrin IX) group per subunit.
CC       {ECO:0000269|PubMed:19625248, ECO:0000269|PubMed:20845962,
CC       ECO:0000269|PubMed:7849045};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=8.0 uM for erythromycin D {ECO:0000269|PubMed:7849045};
CC         Note=kcat is 108 min(-1).;
CC   -!- PATHWAY: Antibiotic biosynthesis; erythromycin biosynthesis.
CC       {ECO:0000269|PubMed:7849045}.
CC   -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:19625248,
CC       ECO:0000269|PubMed:7849045}.
CC   -!- DISRUPTION PHENOTYPE: Cells lacking this gene no longer produce
CC       erythromycin A but accumulate the B and D forms of the antibiotic.
CC       {ECO:0000269|PubMed:8416893}.
CC   -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
DR   EMBL; U82823; AAC45584.1; -; Genomic_DNA.
DR   EMBL; AM420293; CAM00054.1; -; Genomic_DNA.
DR   PIR; B40634; B40634.
DR   RefSeq; WP_009950895.1; NZ_PDBV01000001.1.
DR   PDB; 2JJN; X-ray; 1.59 A; A=1-397.
DR   PDB; 2JJO; X-ray; 1.99 A; A=1-397.
DR   PDB; 2JJP; X-ray; 2.10 A; A=1-397.
DR   PDB; 2WIO; X-ray; 2.00 A; A=2-397.
DR   PDB; 2XFH; X-ray; 1.90 A; A=2-397.
DR   PDB; 3ZKP; X-ray; 2.00 A; A=2-397.
DR   PDBsum; 2JJN; -.
DR   PDBsum; 2JJO; -.
DR   PDBsum; 2JJP; -.
DR   PDBsum; 2WIO; -.
DR   PDBsum; 2XFH; -.
DR   PDBsum; 3ZKP; -.
DR   SMR; P48635; -.
DR   STRING; 405948.SACE_0713; -.
DR   EnsemblBacteria; CAM00054; CAM00054; SACE_0713.
DR   KEGG; sen:SACE_0713; -.
DR   eggNOG; ENOG4106A3M; Bacteria.
DR   eggNOG; COG2124; LUCA.
DR   HOGENOM; HOG000243678; -.
DR   KO; K14370; -.
DR   OMA; PTVREMN; -.
DR   OrthoDB; 816674at2; -.
DR   BioCyc; MetaCyc:MONOMER-17062; -.
DR   BioCyc; SERY405948:SACE_RS03495-MONOMER; -.
DR   BRENDA; 1.14.13.154; 5518.
DR   BRENDA; 1.14.13.185; 5518.
DR   UniPathway; UPA00240; -.
DR   EvolutionaryTrace; P48635; -.
DR   Proteomes; UP000006728; Chromosome.
DR   GO; GO:0020037; F:heme binding; IDA:UniProtKB.
DR   GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR   GO; GO:0004497; F:monooxygenase activity; IMP:UniProtKB.
DR   GO; GO:0050661; F:NADP 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:0033068; P:macrolide biosynthetic 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   SUPFAM; SSF48264; SSF48264; 1.
DR   PROSITE; PS00086; CYTOCHROME_P450; 1.
PE   1: Evidence at protein level;
DR   PRODOM; P48635.
DR   SWISS-2DPAGE; P48635.
KW   3D-structure; Antibiotic biosynthesis; Heme; Iron; Metal-binding;
KW   Monooxygenase; NADP; Oxidoreductase; Reference proteome.
FT   CHAIN           1..397
FT                   /note="Erythromycin C-12 hydroxylase"
FT                   /id="PRO_0000052229"
FT   REGION          74..75
FT                   /note="Substrate binding"
FT   METAL           339
FT                   /note="Iron (heme axial ligand)"
FT   BINDING         81
FT                   /note="Heme"
FT   BINDING         85
FT                   /note="Heme"
FT   BINDING         278
FT                   /note="Substrate"
FT   BINDING         279
FT                   /note="Heme"
FT   BINDING         337
FT                   /note="Heme"
FT   CONFLICT        330
FT                   /note="L -> F (in Ref. 1 and 2; AAC45584)"
FT                   /evidence="ECO:0000305"
FT   STRAND          10..12
FT                   /evidence="ECO:0000244|PDB:2WIO"
FT   HELIX           13..26
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          28..31
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          37..39
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           42..50
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   TURN            52..54
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          55..57
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           59..62
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           73..75
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           80..91
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           94..98
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           101..113
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          117..120
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           121..124
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   TURN            125..127
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           128..138
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           142..144
FT                   /evidence="ECO:0000244|PDB:2XFH"
FT   HELIX           151..157
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           166..192
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           198..204
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          207..209
FT                   /evidence="ECO:0000244|PDB:2WIO"
FT   HELIX           214..245
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           248..255
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           257..259
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           260..270
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          276..283
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          285..287
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          290..292
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          297..301
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           302..306
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   TURN            309..311
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          312..314
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           326..328
FT                   /evidence="ECO:0000244|PDB:2XFH"
FT   HELIX           330..332
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   HELIX           342..360
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          363..365
FT                   /evidence="ECO:0000244|PDB:2JJO"
FT   STRAND          377..383
FT                   /evidence="ECO:0000244|PDB:2JJN"
FT   STRAND          386..388
FT                   /evidence="ECO:0000244|PDB:2JJN"
SQ   SEQUENCE   397 AA;  43725 MW;  0106AF77AFAC86F0 CRC64;
     MTTIDEVPGM ADETALLDWL GTMREKQPVW QDRYGVWHVF RHADVQTVLR DTATFSSDPT
     RVIEGASPTP GMIHEIDPPE HRALRKVVSS AFTPRTISDL EPRIRDVTRS LLADAGESFD
     LVDVLAFPLP VTIVAELLGL PPMDHEQFGD WSGALVDIQM DDPTDPALAE RIADVLNPLT
     AYLKARCAER RADPGDDLIS RLVLAEVDGR ALDDEEAANF STALLLAGHI TTTVLLGNIV
     RTLDEHPAHW DAAAEDPGRI PAIVEEVLRY RPPFPQMQRT TTKATEVAGV PIPADVMVNT
     WVLSANRDSD AHDDPDRFDP SRKSGGAAQL SFGHGVHFCL GAPLARLENR VALEEIIARF
     GRLTVDRDDE RLRHFEQIVL GTRHLPVLAG SSPRQSA
//

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