NCSB1_STRCZ
ID NCSB1_STRCZ Reviewed; 332 AA.
AC Q84HC8;
DT 29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2003, sequence version 1.
DT 03-AUG-2022, entry version 70.
DE RecName: Full=2,7-dihydroxy-5-methyl-1-naphthoate 7-O-methyltransferase {ECO:0000305};
DE EC=2.1.1.303 {ECO:0000269|PubMed:18387946};
DE AltName: Full=Neocarzinostatin O-methyltransferase {ECO:0000305};
DE AltName: Full=Neocarzinostatin biosynthesis protein B1 {ECO:0000305};
GN Name=ncsB1 {ECO:0000303|PubMed:15797213};
OS Streptomyces carzinostaticus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1897;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PATHWAY.
RC STRAIN=ATCC 15944 / E-793 / F-51;
RX PubMed=15797213; DOI=10.1016/j.chembiol.2004.12.013;
RA Liu W., Nonaka K., Nie L., Zhang J., Christenson S.D., Bae J.,
RA Van Lanen S.G., Zazopoulos E., Farnet C.M., Yang C.F., Shen B.;
RT "The neocarzinostatin biosynthetic gene cluster from Streptomyces
RT carzinostaticus ATCC 15944 involving two iterative type I polyketide
RT synthases.";
RL Chem. Biol. 12:293-302(2005).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=ATCC 15944 / E-793 / F-51;
RX PubMed=18387946; DOI=10.1074/jbc.m802206200;
RA Luo Y., Lin S., Zhang J., Cooke H.A., Bruner S.D., Shen B.;
RT "Regiospecific O-methylation of naphthoic acids catalyzed by NcsB1, an O-
RT methyltransferase involved in the biosynthesis of the enediyne antitumor
RT antibiotic neocarzinostatin.";
RL J. Biol. Chem. 283:14694-14702(2008).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.08 ANGSTROMS) IN COMPLEX WITH
RP S-ADENOSYL-L-METHIONINE AND 1,4-DIHYDROXY-2-NAPHTHOATE, AND MUTAGENESIS OF
RP ARG-11 AND TYR-293.
RC STRAIN=ATCC 15944 / E-793 / F-51;
RX PubMed=19702337; DOI=10.1021/bi901257q;
RA Cooke H.A., Guenther E.L., Luo Y., Shen B., Bruner S.D.;
RT "Molecular basis of substrate promiscuity for the SAM-dependent O-
RT methyltransferase NcsB1, involved in the biosynthesis of the enediyne
RT antitumor antibiotic neocarzinostatin.";
RL Biochemistry 48:9590-9598(2009).
CC -!- FUNCTION: S-adenosyl-L-methionine-dependent O-methyltransferase that
CC catalyzes regiospecific methylation at the 7-hydroxy group of 2,7-
CC dihydroxy-5-methyl-1-naphthoate in the biosynthesis of the naphthoate
CC moiety of the neocarzinostatin chromophore. Also recognizes other
CC dihydroxynaphthoate as substrates and catalyzes their regiospecific O-
CC methylation. The carboxylate and its ortho-hydroxy groups of the
CC substrate appear to be crucial for NcsB1 substrate recognition and
CC binding, and O-methylation takes place only at the free hydroxy group
CC of these dihydroxynaphthoic acids. {ECO:0000269|PubMed:18387946}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2,7-dihydroxy-5-methyl-1-naphthoate + S-adenosyl-L-methionine
CC = 2-hydroxy-7-methoxy-5-methyl-1-naphthoate + H(+) + S-adenosyl-L-
CC homocysteine; Xref=Rhea:RHEA:41536, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:78281,
CC ChEBI:CHEBI:78282; EC=2.1.1.303;
CC Evidence={ECO:0000269|PubMed:18387946};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=206 uM for 2,7-dihydroxy-5-methyl-1-naphthoate
CC {ECO:0000269|PubMed:18387946};
CC KM=352 uM for 3,5-dihydroxy-2-naphthoate
CC {ECO:0000269|PubMed:18387946};
CC KM=66 uM for 3,7-dihydroxy-2-naphthoate
CC {ECO:0000269|PubMed:18387946};
CC KM=24 uM for 1,4-dihydroxy-2-naphthoate
CC {ECO:0000269|PubMed:18387946};
CC Note=kcat is 0.69 min(-1) with 2,7-dihydroxy-5-methyl-1-naphthoate as
CC substrate. kcat is 0.27 min(-1) with 3,5-dihydroxy-2-naphthoate as
CC substrate. kcat is 0.030 min(-1) with 3,7-dihydroxy-2-naphthoate as
CC substrate. kcat is 0.0080 min(-1) 1,4-dihydroxy-2-naphthoate with as
CC substrate. {ECO:0000269|PubMed:18387946};
CC pH dependence:
CC Optimum pH is 6.0. {ECO:0000269|PubMed:18387946};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000305|PubMed:15797213}.
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; AY117439; AAM77984.1; -; Genomic_DNA.
DR PDB; 3I53; X-ray; 2.08 A; A/B=1-332.
DR PDB; 3I58; X-ray; 2.69 A; A/B=1-332.
DR PDB; 3I5U; X-ray; 2.60 A; A/B=1-332.
DR PDB; 3I64; X-ray; 3.00 A; A/B=1-332.
DR PDBsum; 3I53; -.
DR PDBsum; 3I58; -.
DR PDBsum; 3I5U; -.
DR PDBsum; 3I64; -.
DR AlphaFoldDB; Q84HC8; -.
DR SMR; Q84HC8; -.
DR KEGG; ag:AAM77984; -.
DR BRENDA; 2.1.1.303; 13527.
DR EvolutionaryTrace; Q84HC8; -.
DR GO; GO:0008171; F:O-methyltransferase activity; IDA:UniProtKB.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IDA:UniProtKB.
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 S-adenosyl-L-methionine; Transferase.
FT CHAIN 1..332
FT /note="2,7-dihydroxy-5-methyl-1-naphthoate 7-O-
FT methyltransferase"
FT /id="PRO_0000430701"
FT ACT_SITE 246
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01020"
FT BINDING 11
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 133
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 153
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 175..179
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 177
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 200
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01020,
FT ECO:0000269|PubMed:19702337"
FT BINDING 227..228
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 242..243
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000269|PubMed:19702337"
FT BINDING 247
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:19702337"
FT MUTAGEN 11
FT /note="R->A,W: Still able to methylate naphthoate. Induces
FT a doubling of KM."
FT /evidence="ECO:0000269|PubMed:19702337"
FT MUTAGEN 11
FT /note="R->K: Increased ability to methylate naphthoate.
FT Increased rate of turnover."
FT /evidence="ECO:0000269|PubMed:19702337"
FT MUTAGEN 293
FT /note="Y->I: Still able to methylate naphthoate. Induces an
FT increase of KM."
FT /evidence="ECO:0000269|PubMed:19702337"
FT HELIX 10..13
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 17..28
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 30..35
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 41..48
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 52..64
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 67..70
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 74..78
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 82..85
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 89..91
FT /evidence="ECO:0007829|PDB:3I64"
FT HELIX 94..97
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 102..106
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 107..112
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 113..119
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 124..128
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 132..138
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 140..158
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 161..163
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 168..170
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 171..176
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 182..190
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 195..200
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 202..214
FT /evidence="ECO:0007829|PDB:3I53"
FT TURN 218..220
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 221..225
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 237..243
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 245..247
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 250..264
FT /evidence="ECO:0007829|PDB:3I53"
FT TURN 265..267
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 269..274
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 284..294
FT /evidence="ECO:0007829|PDB:3I53"
FT HELIX 301..310
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 313..320
FT /evidence="ECO:0007829|PDB:3I53"
FT STRAND 322..331
FT /evidence="ECO:0007829|PDB:3I53"
SQ SEQUENCE 332 AA; 34592 MW; B3DF01BE1EE9D65C CRC64;
MGKRAAHIGL RALADLATPM AVRVAATLRV ADHIAAGHRT AAEIASAAGA HADSLDRLLR
HLVAVGLFTR DGQGVYGLTE FGEQLRDDHA AGKRKWLDMN SAVGRGDLGF VELAHSIRTG
QPAYPVRYGT SFWEDLGSDP VLSASFDTLM SHHLELDYTG IAAKYDWAAL GHVVDVGGGS
GGLLSALLTA HEDLSGTVLD LQGPASAAHR RFLDTGLSGR AQVVVGSFFD PLPAGAGGYV
LSAVLHDWDD LSAVAILRRC AEAAGSGGVV LVIEAVAGDE HAGTGMDLRM LTYFGGKERS
LAELGELAAQ AGLAVRAAHP ISYVSIVEMT AL
