ID EVAA_AMYOR Reviewed; 471 AA. AC O52793; DT 23-MAY-2018, integrated into UniProtKB/Swiss-Prot. DT 01-JUN-1998, sequence version 1. DT 24-JAN-2024, entry version 60. DE RecName: Full=dTDP-4-dehydro-6-deoxy-alpha-D-glucopyranose 2,3-dehydratase {ECO:0000303|PubMed:11035791}; DE EC=4.2.1.159 {ECO:0000269|PubMed:11035791, ECO:0000269|PubMed:23473392}; DE AltName: Full=2,3-dehydratase {ECO:0000303|PubMed:23473392}; GN Name=evaA {ECO:0000303|PubMed:11035791}; GN Synonyms=Orf23 {ECO:0000303|PubMed:11035791}; OS Amycolatopsis orientalis (Nocardia orientalis). OC Bacteria; Actinomycetota; Actinomycetes; Pseudonocardiales; OC Pseudonocardiaceae; Amycolatopsis. OX NCBI_TaxID=31958; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RX PubMed=9545426; DOI=10.1016/s1074-5521(98)90060-6; RA van Wageningen A., Kirkpatrick P., Williams D., Harris B., Kershaw J., RA Lennard N., Jones M., Jones S., Solenberg P.; RT "Sequencing and analysis of genes involved in the biosynthesis of a RT vancomycin group antibiotic."; RL Chem. Biol. 5:155-162(1998). RN [2] RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY. RX PubMed=11035791; DOI=10.1073/pnas.210395097; RA Chen H., Thomas M.G., Hubbard B.K., Losey H.C., Walsh C.T., Burkart M.D.; RT "Deoxysugars in glycopeptide antibiotics: enzymatic synthesis of TDP-L- RT epivancosamine in chloroeremomycin biosynthesis."; RL Proc. Natl. Acad. Sci. U.S.A. 97:11942-11947(2000). RN [3] RP X-RAY CRYSTALLOGRAPHY (1.69 ANGSTROMS) OF MUTANT ALA-381 IN COMPLEX WITH RP SUBSTRATE ANALOGS, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL RP PROPERTIES, MUTAGENESIS OF GLU-139; GLU-190; ASP-353; ARG-381; GLU-404; RP GLU-405 AND GLU-452, DISULFIDE BONDS, SUBUNIT, AND REACTION MECHANISM. RX PubMed=23473392; DOI=10.1021/bi400176n; RA Kubiak R.L., Thoden J.B., Holden H.M.; RT "Structure of EvaA: a paradigm for sugar 2,3-dehydratases."; RL Biochemistry 52:2078-2088(2013). CC -!- FUNCTION: Involved in the biosynthesis of the 2,3,6-trideoxysugar L- CC epivancosamine, the terminal sugar added to the aglycone scaffold of CC chloroeremomycin, a member of the glycopeptide antibiotics vancomycin CC family. Catalyzes the removal of the hydroxyl group at position C-2 of CC the hexose ring of dTDP-4-dehydro-6-deoxy-alpha-D-glucopyranose, and CC the oxidation of the hydroxyl group at position C-3 to form a carbonyl CC functionality. The product of the reaction, dTDP-2,6-dideoxy-D-glycero- CC hex-2-enos-4-ulose, is a highly unstable diketosugar, which CC spontaneously forms dTDP-3,4-didehydro-2,6-dideoxy-alpha-D-glucose. CC {ECO:0000269|PubMed:11035791, ECO:0000269|PubMed:23473392}. CC -!- CATALYTIC ACTIVITY: CC Reaction=dTDP-4-dehydro-6-deoxy-alpha-D-glucose = dTDP-3,4- CC didehydro-2,6-dideoxy-alpha-D-glucose + H2O; Xref=Rhea:RHEA:47972, CC ChEBI:CHEBI:15377, ChEBI:CHEBI:57649, ChEBI:CHEBI:84540; CC EC=4.2.1.159; Evidence={ECO:0000269|PubMed:11035791, CC ECO:0000269|PubMed:23473392}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=56 uM for dTDP-glucose {ECO:0000269|PubMed:23473392}; CC Vmax=4 umol/min/mg enzyme for dTDP-glucose CC {ECO:0000269|PubMed:23473392}; CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000305|PubMed:11035791}. CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:23473392}. CC -!- MISCELLANEOUS: The mutant Ala-381 is used to improve the quality of the CC crystals. {ECO:0000269|PubMed:23473392}. CC -!- MISCELLANEOUS: Two binding sites (pockets A and B) for the dTDP-sugar CC ligands have been identified in each subunit. It seems that pocket A CC represents the active site and pocket B is a vestige of the gene CC duplication event. {ECO:0000269|PubMed:23473392}. CC -!- SIMILARITY: Belongs to the hexose 2,3-dehydratase family. CC {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AJ223998; CAA11763.1; -; Genomic_DNA. DR PIR; T17472; T17472. DR PDB; 4J7G; X-ray; 1.70 A; A/B=1-471. DR PDB; 4J7H; X-ray; 1.69 A; A/B=1-471. DR PDBsum; 4J7G; -. DR PDBsum; 4J7H; -. DR AlphaFoldDB; O52793; -. DR SMR; O52793; -. DR STRING; 31958.SD37_33630; -. DR BioCyc; MetaCyc:MONOMER-18438; -. DR BRENDA; 4.2.1.159; 315. DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW. DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW. DR Gene3D; 3.90.79.40; EvaA sugar 2,3-dehydratase subunit; 3. DR InterPro; IPR005212; EvaA-like. DR InterPro; IPR038153; EvaA-like_sf. DR Pfam; PF03559; Hexose_dehydrat; 2. PE 1: Evidence at protein level; KW 3D-structure; Antibiotic biosynthesis; Disulfide bond; Lyase. FT CHAIN 1..471 FT /note="dTDP-4-dehydro-6-deoxy-alpha-D-glucopyranose 2,3- FT dehydratase" FT /id="PRO_0000444212" FT BINDING 67 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="1" FT /note="from pocket A" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G" FT BINDING 155..159 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="1" FT /note="from pocket A" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G" FT BINDING 193 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="2" FT /note="from pocket B" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G, ECO:0007744|PDB:4J7H" FT BINDING 238 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="1" FT /note="from pocket A" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G" FT BINDING 288 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="2" FT /note="from pocket B" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G, ECO:0007744|PDB:4J7H" FT BINDING 351 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="2" FT /note="from pocket B" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G, ECO:0007744|PDB:4J7H" FT BINDING 367..369 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="2" FT /note="from pocket B" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G, ECO:0007744|PDB:4J7H" FT BINDING 372..373 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="2" FT /note="from pocket B" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G, ECO:0007744|PDB:4J7H" FT BINDING 405..408 FT /ligand="dTDP-4-dehydro-6-deoxy-alpha-D-glucose" FT /ligand_id="ChEBI:CHEBI:57649" FT /ligand_label="1" FT /note="from pocket A" FT /evidence="ECO:0000305|PubMed:23473392, FT ECO:0007744|PDB:4J7G" FT DISULFID 286 FT /note="Interchain" FT /evidence="ECO:0000269|PubMed:23473392, FT ECO:0007744|PDB:4J7G, ECO:0007744|PDB:4J7H" FT MUTAGEN 139 FT /note="E->Q: Loss of dehydratase activity." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 190 FT /note="E->A,Q: Loss of dehydratase activity." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 353 FT /note="D->A: Same dehydratase activity compared to the wild FT type." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 381 FT /note="R->A: The mutation has little effect on the overall FT catalytic efficiency of the enzyme." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 404 FT /note="E->Q: Loss of dehydratase activity." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 405 FT /note="E->A,Q: Loss of dehydratase activity." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 452 FT /note="E->A: Loss of dehydratase activity." FT /evidence="ECO:0000269|PubMed:23473392" FT MUTAGEN 452 FT /note="E->Q: Same dehydratase activity compared to the wild FT type." FT /evidence="ECO:0000269|PubMed:23473392" FT HELIX 21..29 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 38..51 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 55..59 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 61..63 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 65..69 FT /evidence="ECO:0007829|PDB:4J7H" FT TURN 71..73 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 76..78 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 85..94 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 104..112 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 117..126 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 129..138 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 145..149 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 151..154 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 156..158 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 171..173 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 179..181 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 182..189 FT /evidence="ECO:0007829|PDB:4J7H" FT TURN 192..194 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 195..197 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 199..206 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 217..221 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 222..229 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 237..244 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 259..271 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 274..280 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 282..284 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 288..290 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 295..297 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 302..311 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 320..329 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 331..340 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 343..352 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 359..363 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 365..368 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 370..373 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 378..380 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 385..389 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 393..395 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 396..403 FT /evidence="ECO:0007829|PDB:4J7H" FT TURN 406..408 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 409..411 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 413..420 FT /evidence="ECO:0007829|PDB:4J7H" FT STRAND 431..434 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 436..442 FT /evidence="ECO:0007829|PDB:4J7H" FT HELIX 451..465 FT /evidence="ECO:0007829|PDB:4J7H" SQ SEQUENCE 471 AA; 53023 MW; 9D8ED63C2C98CA51 CRC64; MSSFVVPSLT AVRPRDHHDY ADRIALSAAT TDGVQMRTED VRAWIAERRD ANVFHVERIP FADLDQWWFE GVTGNLVHRS GRFFTIEGLH VIEHDGPHGD GPYREWQQPV IRQPEVGILG ILAKEFDGVL HFLMQAKMEP GNPNLVQLSP TVQATRSNYT KAHGGTNVKL IEYFAPPDPE RVIVDVLQAE QGSWFFRKSN RNMIVETVDD VPLWDDFCWL TLGQIAELMH EDETINMNSR SVLSCLPYQD ITPRALFSDV QLLSWFTNER SRHDVRVRRI PLADVCGWKQ GAEEIEHEDG RYFKVLAVAV KGSNREKISW TQPLVESVDL GVVAFLVRKI DGVPHVLVQA RVDGGFLDTV ELAPTVQCTP LNYAHLPAEE RPPFLDLVQN APRSRIRYEA IHSEEGGRFL GVRARYLVID ADEAIDPPPG YAWVTPAQLT ALTRHGHYVN VEARTLLACI NAAAAQPRGG A //