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Protein

Flavin-dependent tryptophan halogenase RebH

Gene

rebH

Organism
Lechevalieria aerocolonigenes (Nocardia aerocolonigenes) (Saccharothrix aerocolonigenes)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Involved in the biosynthesis of the indolocarbazole antitumor agent rebeccamycin. Catalyze the chlorination of tryptophan (Trp) at C7 position to yield 7-chlorotryptophan. The reaction between FADH2, Cl-, and O2 generates the powerful oxidant HOCl, which is presumed to carry out the chlorination reaction. The reaction of HOCl with the active site Lys-79 generates a lysine chloramine, which plays a key role in directing regiospecific chlorination of substrate in this important class of biosynthetic enzymes. It is also able to use bromide ions to generate monobrominated Trp.3 Publications

Catalytic activityi

Tryptophan + FADH2 + chloride + O2 + H+ = 7-chloro-L-tryptophan + FAD + 2 H2O.2 Publications

Cofactori

FAD3 Publications

Kineticsi

Kcat for tryptophan is 1.4 (min-1).

  1. KM=2 µM for tryptophan (with FAD)1 Publication

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Active sitei79 – 7911 Publication
    Binding sitei197 – 1971FAD; via amide nitrogen and carbonyl oxygen2 Publications
    Binding sitei348 – 3481FAD2 Publications
    Binding sitei357 – 3571L-tryptophan; via carbonyl oxygen
    Binding sitei360 – 3601Chloride; via amide nitrogen
    Binding sitei361 – 3611FAD; via amide nitrogen2 Publications
    Binding sitei461 – 4611L-tryptophan
    Binding sitei465 – 4651L-tryptophan; via carbonyl oxygen

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi13 – 164FAD2 Publications
    Nucleotide bindingi39 – 413FAD2 Publications
    Nucleotide bindingi49 – 502FAD2 Publications

    GO - Molecular functioni

    Complete GO annotation...

    Keywords - Molecular functioni

    Oxidoreductase

    Keywords - Ligandi

    FAD, Flavoprotein, Nucleotide-binding

    Enzyme and pathway databases

    BioCyciMetaCyc:MONOMER-15085.
    BRENDAi1.14.14.7. 4340.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Flavin-dependent tryptophan halogenase RebH (EC:1.14.19.9)
    Gene namesi
    Name:rebH
    Synonyms:rbmJ
    OrganismiLechevalieria aerocolonigenes (Nocardia aerocolonigenes) (Saccharothrix aerocolonigenes)
    Taxonomic identifieri68170 [NCBI]
    Taxonomic lineageiBacteriaActinobacteriaPseudonocardialesPseudonocardiaceaeLechevalieria

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi79 – 791K → A: Complete loss of halogenase activity. 1 Publication
    Mutagenesisi79 – 791K → M: Complete loss of halogenase activity. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 530530Flavin-dependent tryptophan halogenase RebHPRO_0000422334Add
    BLAST

    Interactioni

    Subunit structurei

    Homodimer.2 Publications

    Structurei

    Secondary structure

    1
    530
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi7 – 115Combined sources
    Helixi15 – 2713Combined sources
    Turni28 – 303Combined sources
    Beta strandi31 – 388Combined sources
    Helixi54 – 585Combined sources
    Helixi60 – 634Combined sources
    Helixi67 – 737Combined sources
    Beta strandi77 – 793Combined sources
    Beta strandi81 – 899Combined sources
    Beta strandi103 – 11210Combined sources
    Beta strandi116 – 1216Combined sources
    Helixi122 – 13110Combined sources
    Helixi139 – 1435Combined sources
    Helixi146 – 1516Combined sources
    Beta strandi168 – 1714Combined sources
    Helixi173 – 18715Combined sources
    Beta strandi191 – 1944Combined sources
    Beta strandi197 – 2026Combined sources
    Beta strandi208 – 2136Combined sources
    Beta strandi218 – 2203Combined sources
    Beta strandi222 – 2265Combined sources
    Helixi229 – 2313Combined sources
    Helixi233 – 2386Combined sources
    Beta strandi243 – 2453Combined sources
    Turni247 – 2493Combined sources
    Beta strandi254 – 2618Combined sources
    Helixi264 – 2674Combined sources
    Beta strandi271 – 2777Combined sources
    Beta strandi279 – 2879Combined sources
    Beta strandi289 – 29810Combined sources
    Turni300 – 3023Combined sources
    Helixi305 – 31511Combined sources
    Turni320 – 3223Combined sources
    Beta strandi326 – 3294Combined sources
    Beta strandi333 – 3364Combined sources
    Beta strandi338 – 3403Combined sources
    Beta strandi343 – 3453Combined sources
    Turni348 – 3503Combined sources
    Helixi360 – 37314Combined sources
    Helixi382 – 40827Combined sources
    Helixi416 – 4216Combined sources
    Helixi428 – 43811Combined sources
    Beta strandi447 – 4493Combined sources
    Helixi451 – 4566Combined sources
    Helixi458 – 4625Combined sources
    Helixi468 – 47811Combined sources
    Helixi487 – 4915Combined sources
    Helixi493 – 4986Combined sources
    Helixi500 – 51617Combined sources
    Helixi520 – 5278Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    2E4GX-ray2.08A/B1-530[»]
    2O9ZX-ray2.49A/B1-530[»]
    2OA1X-ray2.15A/B1-530[»]
    2OALX-ray2.10A/B1-530[»]
    2OAMX-ray2.30A/B1-530[»]
    4LU6X-ray3.05A/B1-530[»]
    ProteinModelPortaliQ8KHZ8.
    SMRiQ8KHZ8. Positions 1-528.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiQ8KHZ8.

    Family & Domainsi

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni454 – 4552L-tryptophan binding

    Sequence similaritiesi

    Belongs to the RebH family.Curated

    Phylogenomic databases

    KOiK14266.

    Family and domain databases

    Gene3Di3.50.50.60. 3 hits.
    InterProiIPR023753. FAD/NAD-binding_dom.
    IPR006905. Trp_halogenase.
    [Graphical view]
    PfamiPF04820. Trp_halogenase. 1 hit.
    [Graphical view]
    PIRSFiPIRSF011396. Trp_halogenase. 1 hit.
    SUPFAMiSSF51905. SSF51905. 3 hits.

    Sequencei

    Sequence statusi: Complete.

    Q8KHZ8-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MSGKIDKILI VGGGTAGWMA ASYLGKALQG TADITLLQAP DIPTLGVGEA
    60 70 80 90 100
    TIPNLQTAFF DFLGIPEDEW MRECNASYKV AIKFINWRTA GEGTSEAREL
    110 120 130 140 150
    DGGPDHFYHS FGLLKYHEQI PLSHYWFDRS YRGKTVEPFD YACYKEPVIL
    160 170 180 190 200
    DANRSPRRLD GSKVTNYAWH FDAHLVADFL RRFATEKLGV RHVEDRVEHV
    210 220 230 240 250
    QRDANGNIES VRTATGRVFD ADLFVDCSGF RGLLINKAME EPFLDMSDHL
    260 270 280 290 300
    LNDSAVATQV PHDDDANGVE PFTSAIAMKS GWTWKIPMLG RFGTGYVYSS
    310 320 330 340 350
    RFATEDEAVR EFCEMWHLDP ETQPLNRIRF RVGRNRRAWV GNCVSIGTSS
    360 370 380 390 400
    CFVEPLESTG IYFVYAALYQ LVKHFPDKSL NPVLTARFNR EIETMFDDTR
    410 420 430 440 450
    DFIQAHFYFS PRTDTPFWRA NKELRLADGM QEKIDMYRAG MAINAPASDD
    460 470 480 490 500
    AQLYYGNFEE EFRNFWNNSN YYCVLAGLGL VPDAPSPRLA HMPQATESVD
    510 520 530
    EVFGAVKDRQ RNLLETLPSL HEFLRQQHGR
    Length:530
    Mass (Da):60,297
    Last modified:October 1, 2002 - v1
    Checksum:i41825423F03151AB
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AF534707 Genomic DNA. Translation: AAN01216.1.
    AB090952 Genomic DNA. Translation: BAC10682.1.
    AB071405 Genomic DNA. Translation: BAC15758.1.
    AJ414559 Genomic DNA. Translation: CAC93722.1.

    Genome annotation databases

    KEGGiag:BAC10682.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AF534707 Genomic DNA. Translation: AAN01216.1.
    AB090952 Genomic DNA. Translation: BAC10682.1.
    AB071405 Genomic DNA. Translation: BAC15758.1.
    AJ414559 Genomic DNA. Translation: CAC93722.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    2E4GX-ray2.08A/B1-530[»]
    2O9ZX-ray2.49A/B1-530[»]
    2OA1X-ray2.15A/B1-530[»]
    2OALX-ray2.10A/B1-530[»]
    2OAMX-ray2.30A/B1-530[»]
    4LU6X-ray3.05A/B1-530[»]
    ProteinModelPortaliQ8KHZ8.
    SMRiQ8KHZ8. Positions 1-528.
    ModBaseiSearch...
    MobiDBiSearch...

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    KEGGiag:BAC10682.

    Phylogenomic databases

    KOiK14266.

    Enzyme and pathway databases

    BioCyciMetaCyc:MONOMER-15085.
    BRENDAi1.14.14.7. 4340.

    Miscellaneous databases

    EvolutionaryTraceiQ8KHZ8.

    Family and domain databases

    Gene3Di3.50.50.60. 3 hits.
    InterProiIPR023753. FAD/NAD-binding_dom.
    IPR006905. Trp_halogenase.
    [Graphical view]
    PfamiPF04820. Trp_halogenase. 1 hit.
    [Graphical view]
    PIRSFiPIRSF011396. Trp_halogenase. 1 hit.
    SUPFAMiSSF51905. SSF51905. 3 hits.
    ProtoNetiSearch...

    Publicationsi

    1. "The biosynthetic gene cluster for the antitumor rebeccamycin: characterization and generation of indolocarbazole derivatives."
      Sanchez C., Butovich I.A., Brana A.F., Rohr J., Mendez C., Salas J.A.
      Chem. Biol. 9:519-531(2002) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION.
      Strain: ATCC 39243 / DSM 44217.
    2. "Cloning of the staurosporine biosynthetic gene cluster from Streptomyces sp. TP-A0274 and its heterologous expression in Streptomyces lividans."
      Onaka H., Taniguchi S., Igarashi Y., Furumai T.
      J. Antibiot. 55:1063-1071(2002) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    3. "The Biosynthesis of Indolocarbazoles in a Heterologous E. coli Host."
      Hyun C.-G., Bililign T., Liao J., Thorson J.S.
      Submitted (AUG-2002) to the EMBL/GenBank/DDBJ databases
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    4. "Characterization of the biosynthetic gene cluster of rebeccamycin from Lechevalieria aerocolonigenes ATCC 39243."
      Onaka H., Taniguchi S., Igarashi Y., Furumai T.
      Biosci. Biotechnol. Biochem. 67:127-138(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    5. "pTOYAMAcos, pTYM18, and pTYM19, actinomycete-Escherichia coli integrating vectors for heterologous gene expression."
      Onaka H., Taniguchi S., Ikeda H., Igarashi Y., Furumai T.
      J. Antibiot. 56:950-956(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    6. "Molecular analysis of the rebeccamycin L-amino acid oxidase from Lechevalieria aerocolonigenes ATCC 39243."
      Nishizawa T., Aldrich C.C., Sherman D.H.
      J. Bacteriol. 187:2084-2092(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    7. "Biosynthesis of heterocyclic antibiotics in actinomycetes and an approach to synthesize the natural compounds."
      Onaka H.
      Nihon Hosenkin Gakkaishi 20:62-71(2006)
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    8. "Direct formation of chromopyrrolic acid from indole-3-pyruvic acid by StaD, a novel hemoprotein in indolocarbazole biosynthesis."
      Asamizu S., Kato Y., Igarashi Y., Furumai T., Onaka H.
      Tetrahedron Lett. 47:473-475(2006)
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    9. "Robust in vitro activity of RebF and RebH, a two-component reductase/halogenase, generating 7-chlorotryptophan during rebeccamycin biosynthesis."
      Yeh E., Garneau S., Walsh C.T.
      Proc. Natl. Acad. Sci. U.S.A. 102:3960-3965(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR.
    10. "Flavin redox chemistry precedes substrate chlorination during the reaction of the flavin-dependent halogenase RebH."
      Yeh E., Cole L.J., Barr E.W., Bollinger J.M. Jr., Ballou D.P., Walsh C.T.
      Biochemistry 45:7904-7912(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: COFACTOR AND REACTION MECHANISM.
    11. "Chlorination by a long-lived intermediate in the mechanism of flavin-dependent halogenases."
      Yeh E., Blasiak L.C., Koglin A., Drennan C.L., Walsh C.T.
      Biochemistry 46:1284-1292(2007) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.08 ANGSTROMS) IN COMPLEX WITH FAD, CHLORIDE AND TRYPTOPHAN, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE, MUTAGENESIS OF LYS-79, COFACTOR, REACTION MECHANISM, SUBUNIT.
    12. "The structure of flavin-dependent tryptophan 7-halogenase RebH."
      Bitto E., Huang Y., Bingman C.A., Singh S., Thorson J.S., Phillips G.N.
      Proteins 70:289-293(2008) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) IN COMPLEX WITH FAD, CHLORIDE AND TRYPTOPHAN, SUBUNIT.

    Entry informationi

    Entry nameiREBH_NOCAE
    AccessioniPrimary (citable) accession number: Q8KHZ8
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: May 29, 2013
    Last sequence update: October 1, 2002
    Last modified: May 11, 2016
    This is version 56 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure

    Documents

    1. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    2. SIMILARITY comments
      Index of protein domains and families

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.