4OT_BACSU
ID 4OT_BACSU Reviewed; 62 AA.
AC P70994;
DT 10-OCT-2003, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 118.
DE RecName: Full=2-hydroxymuconate tautomerase;
DE EC=5.3.2.6;
DE AltName: Full=(2Z,4E)-2-hydroxyhexa-2,4-dienedioate keto-enol isomerase;
DE AltName: Full=4-oxalocrotonate tautomerase;
DE Short=4-OT;
GN Name=ywhB; OrderedLocusNames=BSU37540;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9353933; DOI=10.1099/00221287-143-10-3313;
RA Presecan E., Moszer I., Boursier L., Cruz Ramos H., De La Fuente V.,
RA Hullo M.-F., Lelong C., Schleich S., Sekowska A., Song B.H., Villani G.,
RA Kunst F., Danchin A., Glaser P.;
RT "The Bacillus subtilis genome from gerBC (311 degrees) to licR (334
RT degrees).";
RL Microbiology 143:3313-3328(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE, MUTAGENESIS OF PRO-2 AND ARG-12,
RP BIOPHYSICOCHEMICAL PROPERTIES, REACTION MECHANISM, AND SUBSTRATE
RP SPECIFICITY.
RX PubMed=17902707; DOI=10.1021/bi701231a;
RA Wang S.C., Johnson W.H. Jr., Czerwinski R.M., Stamps S.L., Whitman C.P.;
RT "Kinetic and stereochemical analysis of YwhB, a 4-oxalocrotonate
RT tautomerase homologue in Bacillus subtilis: mechanistic implications for
RT the YwhB- and 4-oxalocrotonate tautomerase-catalyzed reactions.";
RL Biochemistry 46:11919-11929(2007).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOG, AND
RP SUBUNIT.
RA Hackert M.L., Whitman C.P., Almrud J.J.;
RT "Ywhb binary complex with 2-fluoro-p-hydroxycinnamate.";
RL Submitted (JAN-2007) to the PDB data bank.
CC -!- FUNCTION: Catalyzes both 1,3- and 1,5-keto-enol tautomerization of the
CC diacid 2-hydroxymuconate (2-hydroxy-2,4-hexadienedioate) to produce 2-
CC oxo-4-hexenedioate. This reaction is highly stereoselective and
CC produces a mixture of stereoisomers, where the (3S)-isomer of 2-oxo-4-
CC hexenedioate predominates. Also catalyzes the tautomerization of 2-
CC hydroxymuconate to 2-oxo-3-hexenedioate, however this reaction is
CC slower and occurs after the tautomerization of 2-hydroxymuconate to 2-
CC oxo-4-hexenedioate. Using 2-hydroxy-2,4-pentadienoate,
CC phenylenolpyruvate, (p-hydroxyphenyl)-enolpyruvate and 2-hydroxy-2,4-
CC heptadiene-1,7-dioate, YwhB is a highly efficient 1,3-keto-enol
CC tautomerase, but clearly not a 1,5-keto-enol tautomerase.
CC Tautomerization of the two monoacids 2-hydroxy-2,4-pentadienoate and
CC phenylenolpyruvate produces a mixture of stereoisomers, where the (3R)-
CC isomers predominate. {ECO:0000269|PubMed:17902707}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2Z,4E)-2-hydroxyhexa-2,4-dienedioate = (3E)-2-oxohex-3-
CC enedioate; Xref=Rhea:RHEA:33431, ChEBI:CHEBI:28080,
CC ChEBI:CHEBI:64908; EC=5.3.2.6;
CC Evidence={ECO:0000269|PubMed:17902707};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=76 uM for 2-hydroxymuconate (with 2-oxo-4-hexenedioate as product)
CC {ECO:0000269|PubMed:17902707};
CC KM=160 uM for (p-hydroxyphenyl)-enolpyruvate
CC {ECO:0000269|PubMed:17902707};
CC KM=243 uM for 2-hydroxy-2,4-pentadienoate
CC {ECO:0000269|PubMed:17902707};
CC KM=300 uM for phenylenolpyruvate {ECO:0000269|PubMed:17902707};
CC KM=330 uM for 2-hydroxy-2,4-heptadiene-1,7-dioate
CC {ECO:0000269|PubMed:17902707};
CC KM=940 uM for 2-hydroxymuconate (with 2-oxo-3-hexenedioate as
CC product) {ECO:0000269|PubMed:17902707};
CC Note=kcat is 3 sec(-1) for 2-hydroxy-2,4-heptadiene-1,7-dioate, 4.1
CC sec(-1) for (p-hydroxyphenyl)-enolpyruvate, 10 sec(-1) for 2-
CC hydroxymuconate, 26 sec(-1) for 2-hydroxymuconate, 112 sec(-1) for 2-
CC hydroxy-2,4-pentadienoate, 125 sec(-1) for phenylenolpyruvate.;
CC -!- SUBUNIT: Homohexamer. {ECO:0000305|Ref.4}.
CC -!- MISCELLANEOUS: The mono- and diacids apparently bind in different
CC orientations in the active site of YwhB, but the highly stereoselective
CC nature of the YwhB reaction using a diacid suggests that the biological
CC substrate for YwhB may be a diacid. {ECO:0000305|PubMed:17902707}.
CC -!- SIMILARITY: Belongs to the 4-oxalocrotonate tautomerase family.
CC {ECO:0000305}.
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DR EMBL; Z80360; CAB02512.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB15781.1; -; Genomic_DNA.
DR PIR; C70057; C70057.
DR RefSeq; NP_391634.1; NC_000964.3.
DR RefSeq; WP_003222038.1; NZ_JNCM01000034.1.
DR PDB; 2OP8; X-ray; 2.50 A; A/B=2-62.
DR PDB; 2OPA; X-ray; 2.40 A; A/B=2-62.
DR PDBsum; 2OP8; -.
DR PDBsum; 2OPA; -.
DR AlphaFoldDB; P70994; -.
DR SMR; P70994; -.
DR STRING; 224308.BSU37540; -.
DR PaxDb; P70994; -.
DR PRIDE; P70994; -.
DR EnsemblBacteria; CAB15781; CAB15781; BSU_37540.
DR GeneID; 50135669; -.
DR GeneID; 64305528; -.
DR GeneID; 937098; -.
DR KEGG; bsu:BSU37540; -.
DR PATRIC; fig|224308.179.peg.4065; -.
DR eggNOG; COG1942; Bacteria.
DR InParanoid; P70994; -.
DR OMA; MPIVQVE; -.
DR PhylomeDB; P70994; -.
DR BioCyc; BSUB:BSU37540-MON; -.
DR BioCyc; MetaCyc:BSU37540-MON; -.
DR SABIO-RK; P70994; -.
DR EvolutionaryTrace; P70994; -.
DR PRO; PR:P70994; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0016853; F:isomerase activity; IDA:UniProtKB.
DR GO; GO:0006725; P:cellular aromatic compound metabolic process; IDA:UniProtKB.
DR CDD; cd00491; 4Oxalocrotonate_Tautomerase; 1.
DR Gene3D; 3.30.429.10; -; 1.
DR InterPro; IPR004370; 4-oxalocrotonate_tautomerase.
DR InterPro; IPR014347; Tautomerase/MIF_sf.
DR InterPro; IPR018191; Tautomerase_Pseudo-typ.
DR Pfam; PF01361; Tautomerase; 1.
DR SUPFAM; SSF55331; SSF55331; 1.
DR TIGRFAMs; TIGR00013; taut; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Isomerase; Reference proteome.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000250"
FT CHAIN 2..62
FT /note="2-hydroxymuconate tautomerase"
FT /id="PRO_0000209523"
FT ACT_SITE 2
FT /note="Proton acceptor; via imino nitrogen"
FT /evidence="ECO:0000269|PubMed:17902707"
FT BINDING 9..12
FT /ligand="substrate"
FT MUTAGEN 2
FT /note="P->A: Absence of tautomerase activity using 2-
FT hydroxymuconate. For 2-hydroxy-2,4-pentadienoate and
FT phenylenolpyruvate, a strong decrease of catalytic
FT efficiency and a 2-fold decrease of affinity is observed."
FT /evidence="ECO:0000269|PubMed:17902707"
FT MUTAGEN 12
FT /note="R->A: Absence of tautomerase activity using 2-
FT hydroxymuconate. For 2-hydroxy-2,4-pentadienoate and
FT phenylenolpyruvate, a strong decrease of catalytic
FT efficiency and a 2-fold decrease of affinity is observed."
FT /evidence="ECO:0000269|PubMed:17902707"
FT STRAND 3..10
FT /evidence="ECO:0007829|PDB:2OPA"
FT HELIX 14..32
FT /evidence="ECO:0007829|PDB:2OPA"
FT HELIX 36..38
FT /evidence="ECO:0007829|PDB:2OPA"
FT STRAND 40..46
FT /evidence="ECO:0007829|PDB:2OPA"
FT HELIX 48..50
FT /evidence="ECO:0007829|PDB:2OPA"
FT HELIX 58..60
FT /evidence="ECO:0007829|PDB:2OPA"
SQ SEQUENCE 62 AA; 7145 MW; 1602EDEB61E412EE CRC64;
MPYVTVKMLE GRTDEQKRNL VEKVTEAVKE TTGASEEKIV VFIEEMRKDH YAVAGKRLSD
ME
