ROSA_STRDJ
ID ROSA_STRDJ Reviewed; 347 AA.
AC K4RFM2; E7BBN5;
DT 17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT 09-JAN-2013, sequence version 1.
DT 03-AUG-2022, entry version 49.
DE RecName: Full=8-amino-8-demethylriboflavin N,N-dimethyltransferase {ECO:0000305};
DE EC=2.1.1.343 {ECO:0000269|PubMed:21911488, ECO:0000269|PubMed:26913589};
DE AltName: Full=AF dimethyltransferase {ECO:0000303|PubMed:21911488};
DE AltName: Full=N,N-8-amino-8-demethyl-D-riboflavin dimethyltransferase {ECO:0000303|PubMed:21911488};
GN Name=rosA {ECO:0000303|PubMed:21911488};
GN ORFNames=BN159_8032 {ECO:0000312|EMBL:CCK32410.1};
OS Streptomyces davaonensis (strain DSM 101723 / JCM 4913 / KCC S-0913 / 768).
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1214101;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION BY MASS SPECTROMETRY,
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY, AND
RP DISRUPTION PHENOTYPE.
RC STRAIN=DSM 101723 / JCM 4913 / KCC S-0913 / 768;
RX PubMed=21911488; DOI=10.1074/jbc.m111.292300;
RA Jankowitsch F., Kuehm C., Kellner R., Kalinowski J., Pelzer S.,
RA Macheroux P., Mack M.;
RT "A novel N,N-8-amino-8-demethyl-D-riboflavin dimethyltransferase (RosA)
RT catalyzing the two terminal steps of roseoflavin biosynthesis in
RT Streptomyces davawensis.";
RL J. Biol. Chem. 286:38275-38285(2011).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 101723 / JCM 4913 / KCC S-0913 / 768;
RX PubMed=23043000; DOI=10.1128/jb.01592-12;
RA Jankowitsch F., Schwarz J., Ruckert C., Gust B., Szczepanowski R., Blom J.,
RA Pelzer S., Kalinowski J., Mack M.;
RT "Genome sequence of the bacterium Streptomyces davawensis JCM 4913 and
RT heterologous production of the unique antibiotic roseoflavin.";
RL J. Bacteriol. 194:6818-6827(2012).
RN [3] {ECO:0007744|PDB:4D7K}
RP X-RAY CRYSTALLOGRAPHY (2.22 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=26913589; DOI=10.1111/febs.13690;
RA Tongsook C., Uhl M.K., Jankowitsch F., Mack M., Gruber K., Macheroux P.;
RT "Structural and kinetic studies on RosA, the enzyme catalysing the
RT methylation of 8-demethyl-8-amino-d-riboflavin to the antibiotic
RT roseoflavin.";
RL FEBS J. 283:1531-1549(2016).
CC -!- FUNCTION: Catalyzes the S-adenosyl methionine-dependent conversion of
CC 8-amino-8-demethyl-D-riboflavin (AF) into 8-methylamino-8-demethyl-D-
CC riboflavin (MAF) and roseoflavin (RoF), the last two steps in the
CC biosynthesis of the antibiotic roseoflavin.
CC {ECO:0000269|PubMed:21911488, ECO:0000269|PubMed:26913589}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=8-amino-8-demethylriboflavin + 2 S-adenosyl-L-methionine = 2
CC H(+) + roseoflavin + 2 S-adenosyl-L-homocysteine;
CC Xref=Rhea:RHEA:51944, ChEBI:CHEBI:15378, ChEBI:CHEBI:57856,
CC ChEBI:CHEBI:59789, ChEBI:CHEBI:136518, ChEBI:CHEBI:136521;
CC EC=2.1.1.343; Evidence={ECO:0000269|PubMed:21911488,
CC ECO:0000269|PubMed:26913589};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=57.7 uM for AF {ECO:0000269|PubMed:21911488};
CC KM=4 uM for AF {ECO:0000269|PubMed:26913589};
CC KM=28.6 uM for S-adenosyl-L-methionine {ECO:0000269|PubMed:21911488};
CC KM=70 uM for S-adenosyl-L-methionine {ECO:0000269|PubMed:26913589};
CC Vmax=0.58 umol/min/mg enzyme with AF as substrate
CC {ECO:0000269|PubMed:21911488};
CC Note=kcat is 22.0 min(-1) with AF as substrate. kcat is 14.8 min(-1)
CC with S-adenosyl-L-methionine as substrate.
CC {ECO:0000269|PubMed:21911488};
CC pH dependence:
CC Optimum pH is 8.8. {ECO:0000269|PubMed:21911488};
CC Temperature dependence:
CC Optimum temperature is 52 degrees Celsius.
CC {ECO:0000269|PubMed:21911488};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000269|PubMed:21911488}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:26913589}.
CC -!- DISRUPTION PHENOTYPE: Inactivation of the gene leads to a strain that
CC synthesizes AF but not MAF or RoF. {ECO:0000269|PubMed:21911488}.
CC -!- SIMILARITY: Belongs to the class I-like SAM-binding methyltransferase
CC superfamily. Cation-independent O-methyltransferase family.
CC {ECO:0000255|PROSITE-ProRule:PRU01020}.
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DR EMBL; FR750395; CBY84436.1; -; Genomic_DNA.
DR EMBL; HE971709; CCK32410.1; -; Genomic_DNA.
DR PDB; 4D7K; X-ray; 2.22 A; A/B/C/D/E/F=1-347.
DR PDBsum; 4D7K; -.
DR AlphaFoldDB; K4RFM2; -.
DR SMR; K4RFM2; -.
DR STRING; 1214101.BN159_8032; -.
DR EnsemblBacteria; CCK32410; CCK32410; BN159_8032.
DR KEGG; sdv:BN159_8032; -.
DR PATRIC; fig|1214101.3.peg.8127; -.
DR eggNOG; COG2226; Bacteria.
DR HOGENOM; CLU_005533_12_0_11; -.
DR OMA; PIVGEHE; -.
DR BRENDA; 2.1.1.343; 9909.
DR Proteomes; UP000008043; Chromosome.
DR GO; GO:0008171; F:O-methyltransferase activity; IEA:InterPro.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR Gene3D; 1.10.10.10; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR InterPro; IPR016461; COMT-like.
DR InterPro; IPR001077; O_MeTrfase_dom.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR InterPro; IPR036388; WH-like_DNA-bd_sf.
DR InterPro; IPR036390; WH_DNA-bd_sf.
DR Pfam; PF00891; Methyltransf_2; 1.
DR PIRSF; PIRSF005739; O-mtase; 1.
DR SUPFAM; SSF46785; SSF46785; 1.
DR SUPFAM; SSF53335; SSF53335; 1.
DR PROSITE; PS51683; SAM_OMT_II; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Methyltransferase;
KW Reference proteome; S-adenosyl-L-methionine; Transferase.
FT CHAIN 1..347
FT /note="8-amino-8-demethylriboflavin N,N-
FT dimethyltransferase"
FT /id="PRO_0000449812"
FT BINDING 209
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01020"
FT BINDING 235..237
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01020"
FT HELIX 13..35
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 38..41
FT /evidence="ECO:0007829|PDB:4D7K"
FT TURN 42..44
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 49..56
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 60..72
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 75..79
FT /evidence="ECO:0007829|PDB:4D7K"
FT TURN 80..82
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 83..86
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 88..91
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 102..107
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 111..118
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 120..126
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 131..134
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 135..137
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 139..144
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 146..148
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 149..165
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 167..171
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 176..178
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 180..185
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 191..199
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 200..202
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 204..209
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 211..213
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 214..223
FT /evidence="ECO:0007829|PDB:4D7K"
FT TURN 227..229
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 230..235
FT /evidence="ECO:0007829|PDB:4D7K"
FT TURN 237..239
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 246..252
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 259..270
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 280..285
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 293..305
FT /evidence="ECO:0007829|PDB:4D7K"
FT HELIX 313..322
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 325..333
FT /evidence="ECO:0007829|PDB:4D7K"
FT TURN 334..336
FT /evidence="ECO:0007829|PDB:4D7K"
FT STRAND 337..344
FT /evidence="ECO:0007829|PDB:4D7K"
SQ SEQUENCE 347 AA; 37936 MW; B121C5B441D2E37D CRC64;
MRPEPTEHPE RTAAQRLYQY NVDLKVAFVL YAVAKLHLPD LLADGPRTTA DLAAATGSDP
SRLRRLLRAA AGADALREVP EDSFELAPMG DLLRSGHPRS MRGMTTFFAE PDVLAAYGDL
VESVRTGVPA FQLRHREPLY DFLARPQHKE VRDEFDAAMV EFGQYFADDF LTSFDFGRFT
RFADIGGGRG QFLAGVLTAV PSSTGVLVDG PAVAASAHKF LASQNLTERV EVRIGDFFDV
LPTGCDAYVL RGVLEDWADA DAVRLLVRIR QAMGDAPEAR LLILDSVIGE TGELGKVLDL
DMLVLVEGEH RTRAQWDDLL ARAGFDIVGI HPAGDVWAVI ECRGTAG
