KIJDR_ACTKI
ID KIJDR_ACTKI Reviewed; 332 AA.
AC B3TMR8;
DT 23-MAY-2018, integrated into UniProtKB/Swiss-Prot.
DT 02-SEP-2008, sequence version 1.
DT 03-AUG-2022, entry version 55.
DE RecName: Full=dTDP-3,4-didehydro-2,6-dideoxy-alpha-D-glucose 3-reductase {ECO:0000303|PubMed:21598943};
DE EC=1.1.1.384 {ECO:0000269|PubMed:21598943};
DE AltName: Full=3-ketoreductase {ECO:0000303|PubMed:21598943};
DE AltName: Full=NADPH-dependent C3-ketoreductase {ECO:0000303|PubMed:21598943};
GN ORFNames=KijD10 {ECO:0000312|EMBL:ACB46498.1};
OS Actinomadura kijaniata.
OC Bacteria; Actinobacteria; Streptosporangiales; Thermomonosporaceae;
OC Actinomadura.
OX NCBI_TaxID=46161;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=17985890; DOI=10.1021/ja0744854;
RA Zhang H., White-Phillip J.A., Melancon C.E., Kwon H.-J., Yu W.-L.,
RA Liu H.-W.;
RT "Elucidation of the kijanimicin gene cluster: insights into the
RT biosynthesis of spirotetronate antibiotics and nitrosugars.";
RL J. Am. Chem. Soc. 129:14670-14683(2007).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.71 ANGSTROMS) OF WILD TYPE AND MUTANTS IN COMPLEX
RP WITH NADP AND SUBSTRATE ANALOG, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF LYS-102 AND TYR-186, ACTIVE
RP SITE, PATHWAY, SUBUNIT, AND SUBSTRATE SPECIFICITY.
RX PubMed=21598943; DOI=10.1021/bi200514b;
RA Kubiak R.L., Holden H.M.;
RT "Combined structural and functional investigation of a C-3''-ketoreductase
RT involved in the biosynthesis of dTDP-L-digitoxose.";
RL Biochemistry 50:5905-5917(2011).
CC -!- FUNCTION: Involved in the biosynthesis of L-digitoxose, an unusual
CC dideoxysugar attached to various pharmacologically active natural
CC products, including the antitumor antibiotic tetrocarcin A, and the
CC antibiotics kijanimicin and jadomycin B. Catalyzes the reduction of the
CC C-3 keto moiety of dTDP-3,4-diketo-2,6-dideoxy-alpha-D-glucose to yield
CC dTDP-4-keto-2,6-dideoxy-alpha-D-glucose. Also able to reduce dTDP-3-
CC keto-6-deoxy-D-galactose and dTDP-3-keto-6-deoxy-D-glucose to yield
CC dTDP-fucose and dTDP-quinovose, respectively.
CC {ECO:0000269|PubMed:21598943}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=dTDP-4-dehydro-2,6-dideoxy-alpha-D-glucose + NADP(+) = dTDP-
CC 3,4-didehydro-2,6-dideoxy-alpha-D-glucose + H(+) + NADPH;
CC Xref=Rhea:RHEA:44624, ChEBI:CHEBI:15378, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:84537, ChEBI:CHEBI:84540;
CC EC=1.1.1.384; Evidence={ECO:0000269|PubMed:21598943};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=116.4 uM for dTDP-glucose {ECO:0000269|PubMed:21598943};
CC KM=6.31 mM for NADPH {ECO:0000269|PubMed:21598943};
CC Vmax=0.01 mmol/min/mg enzyme {ECO:0000269|PubMed:21598943};
CC Note=kcat is 6.24 sec(-1) for dTDP-glucose as substrate.
CC {ECO:0000269|PubMed:21598943};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000305|PubMed:21598943}.
CC -!- SUBUNIT: Homotetramer; dimer of dimers. {ECO:0000269|PubMed:21598943}.
CC -!- SIMILARITY: Belongs to the Gfo/Idh/MocA family. {ECO:0000305}.
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DR EMBL; EU301739; ACB46498.1; -; Genomic_DNA.
DR PDB; 3RBV; X-ray; 1.90 A; A=1-332.
DR PDB; 3RC1; X-ray; 1.71 A; A=1-332.
DR PDB; 3RC2; X-ray; 1.80 A; A=1-332.
DR PDB; 3RC7; X-ray; 2.00 A; A=1-332.
DR PDB; 3RC9; X-ray; 1.91 A; A=1-332.
DR PDB; 3RCB; X-ray; 2.49 A; A=1-332.
DR PDBsum; 3RBV; -.
DR PDBsum; 3RC1; -.
DR PDBsum; 3RC2; -.
DR PDBsum; 3RC7; -.
DR PDBsum; 3RC9; -.
DR PDBsum; 3RCB; -.
DR AlphaFoldDB; B3TMR8; -.
DR SMR; B3TMR8; -.
DR BRENDA; 1.1.1.384; 13770.
DR EvolutionaryTrace; B3TMR8; -.
DR GO; GO:0000166; F:nucleotide binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR InterPro; IPR000683; Gfo/Idh/MocA-like_OxRdtase_N.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF01408; GFO_IDH_MocA; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; NADP; Oxidoreductase.
FT CHAIN 1..332
FT /note="dTDP-3,4-didehydro-2,6-dideoxy-alpha-D-glucose 3-
FT reductase"
FT /id="PRO_0000444242"
FT ACT_SITE 102
FT /note="Proton donor"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RC9, ECO:0007744|PDB:3RCB"
FT BINDING 17..23
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RC9, ECO:0007744|PDB:3RCB"
FT BINDING 24
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:21598943,
FT ECO:0007744|PDB:3RC1, ECO:0007744|PDB:3RC2,
FT ECO:0007744|PDB:3RC7, ECO:0007744|PDB:3RC9,
FT ECO:0007744|PDB:3RCB"
FT BINDING 42..43
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RC9, ECO:0007744|PDB:3RCB"
FT BINDING 63
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RC9, ECO:0007744|PDB:3RCB"
FT BINDING 79
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RCB"
FT BINDING 84
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RCB"
FT BINDING 170
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RC1, ECO:0007744|PDB:3RC7"
FT BINDING 182
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21598943,
FT ECO:0007744|PDB:3RBV, ECO:0007744|PDB:3RC1,
FT ECO:0007744|PDB:3RC2, ECO:0007744|PDB:3RC7,
FT ECO:0007744|PDB:3RC9, ECO:0007744|PDB:3RCB"
FT BINDING 240
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:21598943,
FT ECO:0007744|PDB:3RC1, ECO:0007744|PDB:3RC2,
FT ECO:0007744|PDB:3RC7, ECO:0007744|PDB:3RC9,
FT ECO:0007744|PDB:3RCB"
FT BINDING 260
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:21598943,
FT ECO:0007744|PDB:3RC1, ECO:0007744|PDB:3RC2,
FT ECO:0007744|PDB:3RC7, ECO:0007744|PDB:3RC9,
FT ECO:0007744|PDB:3RCB"
FT MUTAGEN 102
FT /note="K->A,M,Q: Loss of reductase activity."
FT /evidence="ECO:0000269|PubMed:21598943"
FT MUTAGEN 102
FT /note="K->E: Retains some activity, but the catalytic
FT efficiency is strongly reduced."
FT /evidence="ECO:0000269|PubMed:21598943"
FT MUTAGEN 186
FT /note="Y->F: Same affinity for dTDP-glucose and NADPH
FT compared to the wild-type. Small reduction of the catalytic
FT efficiency resulting from the conformational flexibility of
FT the nicotinamide ring."
FT /evidence="ECO:0000269|PubMed:21598943"
FT STRAND 10..16
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 19..23
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 25..31
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 35..44
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 45..55
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 57..62
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 63..67
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 73..77
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 81..83
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 84..93
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 97..104
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 108..120
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 125..128
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 130..133
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 136..145
FT /evidence="ECO:0007829|PDB:3RC1"
FT TURN 146..149
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 151..160
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 169..171
FT /evidence="ECO:0007829|PDB:3RC1"
FT TURN 173..176
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 178..182
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 185..195
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 200..209
FT /evidence="ECO:0007829|PDB:3RC1"
FT TURN 210..213
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 214..223
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 229..238
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 241..250
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 252..257
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 268..273
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 276..281
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 287..301
FT /evidence="ECO:0007829|PDB:3RC1"
FT HELIX 306..325
FT /evidence="ECO:0007829|PDB:3RC1"
FT STRAND 326..331
FT /evidence="ECO:0007829|PDB:3RC1"
SQ SEQUENCE 332 AA; 37033 MW; 5122363DF9A8BA51 CRC64;
MENPANANPI RVGVIGCADI AWRRALPALE AEPLTEVTAI ASRRWDRAKR FTERFGGEPV
EGYPALLERD DVDAVYVPLP AVLHAEWIDR ALRAGKHVLA EKPLTTDRPQ AERLFAVARE
RGLLLMENFM FLHHPQHRQV ADMLDEGVIG EIRSFAASFT IPPKPQGDIR YQADVGGGAL
LDIGVYPIRA AGLFLGADLE FVGAVLRHER DRDVVVGGNA LLTTRQGVTA QLTFGMEHAY
TNNYEFRGST GRLWMNRVFT PPATYQPVVH IERQDHAEQF VLPAHDQFAK SIRAFAQAVL
SGEHPREWSE DSLRQASLVD AVRTGARDIY FP
