REBH_LENAE
ID REBH_LENAE Reviewed; 530 AA.
AC Q8KHZ8;
DT 29-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2002, sequence version 1.
DT 03-AUG-2022, entry version 79.
DE RecName: Full=Flavin-dependent tryptophan halogenase RebH;
DE EC=1.14.19.9;
GN Name=rebH; Synonyms=rbmJ;
OS Lentzea aerocolonigenes (Lechevalieria aerocolonigenes) (Saccharothrix
OS aerocolonigenes).
OC Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae; Lentzea.
OX NCBI_TaxID=68170;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=ATCC 39243 / DSM 44217 / BCRC 13729 / KCTC 9384;
RX PubMed=11983340; DOI=10.1016/s1074-5521(02)00126-6;
RA Sanchez C., Butovich I.A., Brana A.F., Rohr J., Mendez C., Salas J.A.;
RT "The biosynthetic gene cluster for the antitumor rebeccamycin:
RT characterization and generation of indolocarbazole derivatives.";
RL Chem. Biol. 9:519-531(2002).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=12617516; DOI=10.7164/antibiotics.55.1063;
RA Onaka H., Taniguchi S., Igarashi Y., Furumai T.;
RT "Cloning of the staurosporine biosynthetic gene cluster from Streptomyces
RT sp. TP-A0274 and its heterologous expression in Streptomyces lividans.";
RL J. Antibiot. 55:1063-1071(2002).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Hyun C.-G., Bililign T., Liao J., Thorson J.S.;
RT "The Biosynthesis of Indolocarbazoles in a Heterologous E. coli Host.";
RL Submitted (AUG-2002) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=12619684; DOI=10.1271/bbb.67.127;
RA Onaka H., Taniguchi S., Igarashi Y., Furumai T.;
RT "Characterization of the biosynthetic gene cluster of rebeccamycin from
RT Lechevalieria aerocolonigenes ATCC 39243.";
RL Biosci. Biotechnol. Biochem. 67:127-138(2003).
RN [5]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=14763561; DOI=10.7164/antibiotics.56.950;
RA Onaka H., Taniguchi S., Ikeda H., Igarashi Y., Furumai T.;
RT "pTOYAMAcos, pTYM18, and pTYM19, actinomycete-Escherichia coli integrating
RT vectors for heterologous gene expression.";
RL J. Antibiot. 56:950-956(2003).
RN [6]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=15743957; DOI=10.1128/jb.187.6.2084-2092.2005;
RA Nishizawa T., Aldrich C.C., Sherman D.H.;
RT "Molecular analysis of the rebeccamycin L-amino acid oxidase from
RT Lechevalieria aerocolonigenes ATCC 39243.";
RL J. Bacteriol. 187:2084-2092(2005).
RN [7]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Onaka H.;
RT "Biosynthesis of heterocyclic antibiotics in actinomycetes and an approach
RT to synthesize the natural compounds.";
RL Nihon Hosenkin Gakkaishi 20:62-71(2006).
RN [8]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Asamizu S., Kato Y., Igarashi Y., Furumai T., Onaka H.;
RT "Direct formation of chromopyrrolic acid from indole-3-pyruvic acid by
RT StaD, a novel hemoprotein in indolocarbazole biosynthesis.";
RL Tetrahedron Lett. 47:473-475(2006).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=15743914; DOI=10.1073/pnas.0500755102;
RA Yeh E., Garneau S., Walsh C.T.;
RT "Robust in vitro activity of RebF and RebH, a two-component
RT reductase/halogenase, generating 7-chlorotryptophan during rebeccamycin
RT biosynthesis.";
RL Proc. Natl. Acad. Sci. U.S.A. 102:3960-3965(2005).
RN [10]
RP REACTION MECHANISM.
RX PubMed=16784243; DOI=10.1021/bi060607d;
RA Yeh E., Cole L.J., Barr E.W., Bollinger J.M. Jr., Ballou D.P., Walsh C.T.;
RT "Flavin redox chemistry precedes substrate chlorination during the reaction
RT of the flavin-dependent halogenase RebH.";
RL Biochemistry 45:7904-7912(2006).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (2.08 ANGSTROMS) IN COMPLEX WITH FAD, CHLORIDE AND
RP TRYPTOPHAN, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE, MUTAGENESIS OF
RP LYS-79, REACTION MECHANISM, AND SUBUNIT.
RX PubMed=17260957; DOI=10.1021/bi0621213;
RA Yeh E., Blasiak L.C., Koglin A., Drennan C.L., Walsh C.T.;
RT "Chlorination by a long-lived intermediate in the mechanism of flavin-
RT dependent halogenases.";
RL Biochemistry 46:1284-1292(2007).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) IN COMPLEX WITH FAD, CHLORIDE AND
RP TRYPTOPHAN, AND SUBUNIT.
RX PubMed=17876823; DOI=10.1002/prot.21627;
RA Bitto E., Huang Y., Bingman C.A., Singh S., Thorson J.S., Phillips G.N.;
RT "The structure of flavin-dependent tryptophan 7-halogenase RebH.";
RL Proteins 70:289-293(2008).
CC -!- FUNCTION: Involved in the biosynthesis of the indolocarbazole antitumor
CC agent rebeccamycin. Catalyzes the chlorination of tryptophan (Trp) at
CC C7 position to yield 7-chlorotryptophan. The reaction between FADH2,
CC Cl-, and O2 generates the powerful oxidant HOCl, which is presumed to
CC carry out the chlorination reaction. The reaction of HOCl with the
CC active site Lys-79 generates a lysine chloramine, which plays a key
CC role in directing regiospecific chlorination of substrate in this
CC important class of biosynthetic enzymes. It is also able to use bromide
CC ions to generate monobrominated Trp. {ECO:0000269|PubMed:11983340,
CC ECO:0000269|PubMed:15743914, ECO:0000269|PubMed:17260957}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=chloride + FADH2 + L-tryptophan + O2 = 7-chloro-L-tryptophan +
CC FAD + 2 H2O; Xref=Rhea:RHEA:26494, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17996, ChEBI:CHEBI:57692,
CC ChEBI:CHEBI:57912, ChEBI:CHEBI:58307, ChEBI:CHEBI:58713;
CC EC=1.14.19.9; Evidence={ECO:0000269|PubMed:15743914,
CC ECO:0000269|PubMed:17260957};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2 uM for tryptophan (with FAD) {ECO:0000269|PubMed:15743914};
CC Note=kcat for tryptophan is 1.4 min(-1).;
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:17260957,
CC ECO:0000269|PubMed:17876823}.
CC -!- SIMILARITY: Belongs to the flavin-dependent halogenase family.
CC Bacterial tryptophan halogenase subfamily. {ECO:0000305}.
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DR EMBL; AF534707; AAN01216.1; -; Genomic_DNA.
DR EMBL; AB090952; BAC10682.1; -; Genomic_DNA.
DR EMBL; AB071405; BAC15758.1; -; Genomic_DNA.
DR EMBL; AJ414559; CAC93722.1; -; Genomic_DNA.
DR PDB; 2E4G; X-ray; 2.08 A; A/B=1-530.
DR PDB; 2O9Z; X-ray; 2.49 A; A/B=1-530.
DR PDB; 2OA1; X-ray; 2.15 A; A/B=1-530.
DR PDB; 2OAL; X-ray; 2.10 A; A/B=1-530.
DR PDB; 2OAM; X-ray; 2.30 A; A/B=1-530.
DR PDB; 4LU6; X-ray; 3.05 A; A/B=1-530.
DR PDB; 6P00; X-ray; 2.25 A; A/B=1-530.
DR PDB; 6P2V; X-ray; 2.55 A; A/B=1-530.
DR PDB; 7JU0; X-ray; 2.60 A; A/B=1-530.
DR PDBsum; 2E4G; -.
DR PDBsum; 2O9Z; -.
DR PDBsum; 2OA1; -.
DR PDBsum; 2OAL; -.
DR PDBsum; 2OAM; -.
DR PDBsum; 4LU6; -.
DR PDBsum; 6P00; -.
DR PDBsum; 6P2V; -.
DR PDBsum; 7JU0; -.
DR AlphaFoldDB; Q8KHZ8; -.
DR SMR; Q8KHZ8; -.
DR KEGG; ag:BAC10682; -.
DR BioCyc; MetaCyc:MON-15085; -.
DR BRENDA; 1.14.19.9; 4340.
DR EvolutionaryTrace; Q8KHZ8; -.
DR GO; GO:0004497; F:monooxygenase activity; IEA:InterPro.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR006905; Flavin_halogenase.
DR InterPro; IPR033856; Trp_halogen.
DR Pfam; PF04820; Trp_halogenase; 1.
DR PIRSF; PIRSF011396; Trp_halogenase; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; FAD; Flavoprotein; Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..530
FT /note="Flavin-dependent tryptophan halogenase RebH"
FT /id="PRO_0000422334"
FT ACT_SITE 79
FT /evidence="ECO:0000269|PubMed:17260957"
FT BINDING 13..16
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17260957,
FT ECO:0000269|PubMed:17876823"
FT BINDING 39..50
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:P95480"
FT BINDING 39..41
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17260957,
FT ECO:0000269|PubMed:17876823"
FT BINDING 49..50
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17260957,
FT ECO:0000269|PubMed:17876823"
FT BINDING 197
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17260957,
FT ECO:0000269|PubMed:17876823"
FT BINDING 348
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17260957,
FT ECO:0000269|PubMed:17876823"
FT BINDING 357
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 359..360
FT /ligand="chloride"
FT /ligand_id="ChEBI:CHEBI:17996"
FT /evidence="ECO:0000250|UniProtKB:P95480"
FT BINDING 360
FT /ligand="chloride"
FT /ligand_id="ChEBI:CHEBI:17996"
FT BINDING 361
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17260957,
FT ECO:0000269|PubMed:17876823"
FT BINDING 454..455
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 461
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 465
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT MUTAGEN 79
FT /note="K->A: Complete loss of halogenase activity."
FT /evidence="ECO:0000269|PubMed:17260957"
FT MUTAGEN 79
FT /note="K->M: Complete loss of halogenase activity."
FT /evidence="ECO:0000269|PubMed:17260957"
FT STRAND 7..11
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 15..27
FT /evidence="ECO:0007829|PDB:2E4G"
FT TURN 28..30
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 31..38
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 54..58
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 60..63
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 67..73
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 77..79
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 81..89
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 103..112
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 116..121
FT /evidence="ECO:0007829|PDB:4LU6"
FT HELIX 122..131
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 139..143
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 146..151
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 168..171
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 173..187
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 191..194
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 197..202
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 208..213
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 218..220
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 222..226
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 229..231
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 233..238
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 243..245
FT /evidence="ECO:0007829|PDB:2E4G"
FT TURN 247..249
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 254..261
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 264..267
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 271..277
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 279..287
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 289..298
FT /evidence="ECO:0007829|PDB:2E4G"
FT TURN 300..302
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 305..315
FT /evidence="ECO:0007829|PDB:2E4G"
FT TURN 320..322
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 326..329
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 333..336
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 338..340
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 343..345
FT /evidence="ECO:0007829|PDB:2E4G"
FT TURN 348..350
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 360..373
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 382..408
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 416..421
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 428..438
FT /evidence="ECO:0007829|PDB:2E4G"
FT STRAND 447..449
FT /evidence="ECO:0007829|PDB:2OAL"
FT HELIX 451..456
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 458..462
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 468..478
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 487..491
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 493..498
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 500..516
FT /evidence="ECO:0007829|PDB:2E4G"
FT HELIX 520..527
FT /evidence="ECO:0007829|PDB:2E4G"
SQ SEQUENCE 530 AA; 60297 MW; 41825423F03151AB CRC64;
MSGKIDKILI VGGGTAGWMA ASYLGKALQG TADITLLQAP DIPTLGVGEA TIPNLQTAFF
DFLGIPEDEW MRECNASYKV AIKFINWRTA GEGTSEAREL DGGPDHFYHS FGLLKYHEQI
PLSHYWFDRS YRGKTVEPFD YACYKEPVIL DANRSPRRLD GSKVTNYAWH FDAHLVADFL
RRFATEKLGV RHVEDRVEHV QRDANGNIES VRTATGRVFD ADLFVDCSGF RGLLINKAME
EPFLDMSDHL LNDSAVATQV PHDDDANGVE PFTSAIAMKS GWTWKIPMLG RFGTGYVYSS
RFATEDEAVR EFCEMWHLDP ETQPLNRIRF RVGRNRRAWV GNCVSIGTSS CFVEPLESTG
IYFVYAALYQ LVKHFPDKSL NPVLTARFNR EIETMFDDTR DFIQAHFYFS PRTDTPFWRA
NKELRLADGM QEKIDMYRAG MAINAPASDD AQLYYGNFEE EFRNFWNNSN YYCVLAGLGL
VPDAPSPRLA HMPQATESVD EVFGAVKDRQ RNLLETLPSL HEFLRQQHGR
