PANE_ECOLI
ID PANE_ECOLI Reviewed; 303 AA.
AC P0A9J4; P77728; Q2MC01; Q46947;
DT 19-JUL-2005, integrated into UniProtKB/Swiss-Prot.
DT 19-JUL-2005, sequence version 1.
DT 03-AUG-2022, entry version 137.
DE RecName: Full=2-dehydropantoate 2-reductase {ECO:0000305};
DE EC=1.1.1.169 {ECO:0000269|PubMed:10736170, ECO:0000269|PubMed:11123955};
DE AltName: Full=Ketopantoate reductase {ECO:0000303|PubMed:10736170};
DE Short=KPA reductase;
DE Short=KPR {ECO:0000303|PubMed:10736170};
GN Name=panE; Synonyms=apbA; OrderedLocusNames=b0425, JW0415;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RA Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT "Sequence of minutes 4-25 of Escherichia coli.";
RL Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 180-303.
RC STRAIN=K12;
RA Backstrom A.D.;
RT "The biosynthesis of thiamin in Escherichia coli K-12: structural genes for
RT thiamin biosynthetic enzymes (thiCEFGH and thiJ) and function of the thiE
RT gene product (thiamin phosphate synthase [E.C. 2.5.1.3]).";
RL Thesis (1996), Cornell University, United States.
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RC STRAIN=K12;
RX PubMed=10736170; DOI=10.1021/bi992676g;
RA Zheng R., Blanchard J.S.;
RT "Kinetic and mechanistic analysis of the E. coli panE-encoded ketopantoate
RT reductase.";
RL Biochemistry 39:3708-3717(2000).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITE,
RP AND MUTAGENESIS OF LYS-176 AND GLU-256.
RX PubMed=11123955; DOI=10.1021/bi002134v;
RA Zheng R., Blanchard J.S.;
RT "Identification of active site residues in E. coli ketopantoate reductase
RT by mutagenesis and chemical rescue.";
RL Biochemistry 39:16244-16251(2000).
RN [7]
RP LIGAND BINDING.
RX PubMed=16042595; DOI=10.1042/bst0330767;
RA Ciulli A., Abell C.;
RT "Biophysical tools to monitor enzyme-ligand interactions of enzymes
RT involved in vitamin biosynthesis.";
RL Biochem. Soc. Trans. 33:767-771(2005).
RN [8]
RP LIGAND BINDING, AND MUTAGENESIS OF ARG-31 AND ASN-98.
RX PubMed=16884311; DOI=10.1021/jm060490r;
RA Ciulli A., Williams G., Smith A.G., Blundell T.L., Abell C.;
RT "Probing hot spots at protein-ligand binding sites: a fragment-based
RT approach using biophysical methods.";
RL J. Med. Chem. 49:4992-5000(2006).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS), PROTEIN SEQUENCE OF 1-9, MASS
RP SPECTROMETRY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=11724562; DOI=10.1021/bi011020w;
RA Matak-Vinkovic D., Vinkovic M., Saldanha S.A., Ashurst J.L., von Delft F.,
RA Inoue T., Miguel R.N., Smith A.G., Blundell T.L., Abell C.;
RT "Crystal structure of Escherichia coli ketopantoate reductase at 1.7 A
RT resolution and insight into the enzyme mechanism.";
RL Biochemistry 40:14493-14500(2001).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (2.09 ANGSTROMS) IN COMPLEX WITH NADP.
RX PubMed=15966718; DOI=10.1021/bi0502036;
RA Lobley C.M.C., Ciulli A., Whitney H.M., Williams G., Smith A.G., Abell C.,
RA Blundell T.L.;
RT "The crystal structure of Escherichia coli ketopantoate reductase with
RT NADP+ bound.";
RL Biochemistry 44:8930-8939(2005).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEX WITH NADP ANALOG, AND
RP MUTAGENESIS OF ASN-98; LYS-176 AND GLU-256.
RX PubMed=17242510; DOI=10.1107/s0907444906044465;
RA Ciulli A., Lobley C.M.C., Tuck K.L., Smith A.G., Blundell T.L., Abell C.;
RT "pH-tuneable binding of 2'-phospho-ADP-ribose to ketopantoate reductase: a
RT structural and calorimetric study.";
RL Acta Crystallogr. D 63:171-178(2007).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND NADP,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF LYS-72 AND SER-244.
RX PubMed=17229734; DOI=10.1074/jbc.m611171200;
RA Ciulli A., Chirgadze D.Y., Smith A.G., Blundell T.L., Abell C.;
RT "Crystal structure of Escherichia coli ketopantoate reductase in a ternary
RT complex with NADP+ and pantoate bound: substrate recognition,
RT conformational change, and cooperativity.";
RL J. Biol. Chem. 282:8487-8497(2007).
CC -!- FUNCTION: Catalyzes the NADPH-dependent reduction of ketopantoate into
CC pantoic acid. {ECO:0000269|PubMed:10736170,
CC ECO:0000269|PubMed:11123955}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(R)-pantoate + NADP(+) = 2-dehydropantoate + H(+) + NADPH;
CC Xref=Rhea:RHEA:16233, ChEBI:CHEBI:11561, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15980, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC EC=1.1.1.169; Evidence={ECO:0000269|PubMed:10736170,
CC ECO:0000269|PubMed:11123955};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=30 uM for ketopantoate {ECO:0000269|PubMed:11724562,
CC ECO:0000269|PubMed:17229734};
CC KM=120 uM for ketopantoate {ECO:0000269|PubMed:10736170,
CC ECO:0000269|PubMed:11123955};
CC KM=7.3 uM for NADPH {ECO:0000269|PubMed:11724562,
CC ECO:0000269|PubMed:17229734};
CC KM=4 uM for NADPH {ECO:0000269|PubMed:10736170,
CC ECO:0000269|PubMed:11123955};
CC KM=260 uM for pantoate {ECO:0000269|PubMed:10736170};
CC KM=7 uM for NADP(+) {ECO:0000269|PubMed:10736170};
CC Vmax=13.8 umol/min/mg enzyme for the reduction of ketopantoate
CC {ECO:0000269|PubMed:10736170};
CC Vmax=0.90 umol/min/mg enzyme for the oxidation of pantoate
CC {ECO:0000269|PubMed:10736170};
CC -!- PATHWAY: Cofactor biosynthesis; (R)-pantothenate biosynthesis; (R)-
CC pantoate from 3-methyl-2-oxobutanoate: step 2/2.
CC {ECO:0000305|PubMed:10736170}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:11724562,
CC ECO:0000269|PubMed:15966718, ECO:0000269|PubMed:17229734,
CC ECO:0000269|PubMed:17242510}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- MASS SPECTROMETRY: Mass=33976; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:11724562};
CC -!- SIMILARITY: Belongs to the ketopantoate reductase family.
CC {ECO:0000305}.
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DR EMBL; U82664; AAB40181.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73528.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76205.1; -; Genomic_DNA.
DR EMBL; U34923; AAA82703.1; -; Genomic_DNA.
DR PIR; A64772; A64772.
DR RefSeq; NP_414959.1; NC_000913.3.
DR RefSeq; WP_000705865.1; NZ_SSZK01000009.1.
DR PDB; 1KS9; X-ray; 1.70 A; A=1-291.
DR PDB; 1YJQ; X-ray; 2.09 A; A=1-303.
DR PDB; 1YON; X-ray; 1.95 A; A=1-303.
DR PDB; 2OFP; X-ray; 2.30 A; A/B=1-303.
DR PDBsum; 1KS9; -.
DR PDBsum; 1YJQ; -.
DR PDBsum; 1YON; -.
DR PDBsum; 2OFP; -.
DR AlphaFoldDB; P0A9J4; -.
DR BMRB; P0A9J4; -.
DR SMR; P0A9J4; -.
DR BioGRID; 4262981; 324.
DR BioGRID; 849454; 1.
DR IntAct; P0A9J4; 5.
DR STRING; 511145.b0425; -.
DR BindingDB; P0A9J4; -.
DR ChEMBL; CHEMBL4855; -.
DR DrugCentral; P0A9J4; -.
DR jPOST; P0A9J4; -.
DR PaxDb; P0A9J4; -.
DR PRIDE; P0A9J4; -.
DR EnsemblBacteria; AAC73528; AAC73528; b0425.
DR EnsemblBacteria; BAE76205; BAE76205; BAE76205.
DR GeneID; 945065; -.
DR KEGG; ecj:JW0415; -.
DR KEGG; eco:b0425; -.
DR PATRIC; fig|1411691.4.peg.1852; -.
DR EchoBASE; EB3056; -.
DR eggNOG; COG1893; Bacteria.
DR HOGENOM; CLU_031468_0_1_6; -.
DR InParanoid; P0A9J4; -.
DR OMA; NYSSMYQ; -.
DR PhylomeDB; P0A9J4; -.
DR BioCyc; EcoCyc:2-DEHYDROPANTOATE-REDUCT-MON; -.
DR BioCyc; MetaCyc:2-DEHYDROPANTOATE-REDUCT-MON; -.
DR BRENDA; 1.1.1.169; 2026.
DR SABIO-RK; P0A9J4; -.
DR UniPathway; UPA00028; UER00004.
DR EvolutionaryTrace; P0A9J4; -.
DR PRO; PR:P0A9J4; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0008677; F:2-dehydropantoate 2-reductase activity; IDA:EcoCyc.
DR GO; GO:0050661; F:NADP binding; IDA:EcoCyc.
DR GO; GO:0015940; P:pantothenate biosynthetic process; IMP:EcoCyc.
DR Gene3D; 1.10.1040.10; -; 1.
DR InterPro; IPR008927; 6-PGluconate_DH-like_C_sf.
DR InterPro; IPR013328; 6PGD_dom2.
DR InterPro; IPR003710; ApbA.
DR InterPro; IPR013752; KPA_reductase.
DR InterPro; IPR013332; KPR_N.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF02558; ApbA; 1.
DR Pfam; PF08546; ApbA_C; 1.
DR SUPFAM; SSF48179; SSF48179; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR00745; apbA_panE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Direct protein sequencing; NADP; Oxidoreductase;
KW Pantothenate biosynthesis; Reference proteome.
FT CHAIN 1..303
FT /note="2-dehydropantoate 2-reductase"
FT /id="PRO_0000157301"
FT ACT_SITE 176
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:11123955"
FT BINDING 7..12
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15966718,
FT ECO:0000269|PubMed:17229734"
FT BINDING 31
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15966718,
FT ECO:0000269|PubMed:17229734"
FT BINDING 98
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15966718,
FT ECO:0000269|PubMed:17229734"
FT BINDING 98
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:17229734"
FT BINDING 122
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15966718,
FT ECO:0000269|PubMed:17229734"
FT BINDING 180
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:17229734"
FT BINDING 184
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:17229734"
FT BINDING 194
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:17229734"
FT BINDING 241
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:17229734"
FT BINDING 244
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:17229734"
FT BINDING 256
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:15966718,
FT ECO:0000269|PubMed:17229734"
FT MUTAGEN 31
FT /note="R->A: Increases KM for NADP 4-fold."
FT /evidence="ECO:0000269|PubMed:16884311"
FT MUTAGEN 72
FT /note="K->A: Increases KM for NADP 10-fold."
FT /evidence="ECO:0000269|PubMed:17229734"
FT MUTAGEN 98
FT /note="N->A: Increases KM for ketopantoate 140-fold.
FT Strongly decreases catalytic activity."
FT /evidence="ECO:0000269|PubMed:16884311,
FT ECO:0000269|PubMed:17242510"
FT MUTAGEN 176
FT /note="K->A: Increases KM for ketopantoate 1400-fold.
FT Decreases Vmax 230-fold. Loss of activity; when associated
FT with A-256."
FT /evidence="ECO:0000269|PubMed:11123955,
FT ECO:0000269|PubMed:17242510"
FT MUTAGEN 244
FT /note="S->A: Increases KM for ketopantoate 230-fold."
FT /evidence="ECO:0000269|PubMed:17229734"
FT MUTAGEN 256
FT /note="E->A: Increases KM for ketopantoate 1100-fold.
FT Decreases Vmax 40-fold. Loss of activity; when associated
FT with A-176."
FT /evidence="ECO:0000269|PubMed:11123955,
FT ECO:0000269|PubMed:17242510"
FT STRAND 2..6
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 10..21
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 25..29
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 35..42
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 44..46
FT /evidence="ECO:0007829|PDB:2OFP"
FT STRAND 48..56
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 58..62
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 65..69
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 73..75
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 76..84
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 93..96
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 98..100
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 104..106
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 113..118
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 121..125
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 128..133
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 137..143
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 144..146
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 151..157
FT /evidence="ECO:0007829|PDB:1KS9"
FT STRAND 163..165
FT /evidence="ECO:0007829|PDB:1KS9"
FT TURN 167..169
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 170..189
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 194..199
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 201..218
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 224..237
FT /evidence="ECO:0007829|PDB:1KS9"
FT TURN 238..240
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 244..250
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 257..259
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 261..271
FT /evidence="ECO:0007829|PDB:1KS9"
FT HELIX 276..290
FT /evidence="ECO:0007829|PDB:1KS9"
SQ SEQUENCE 303 AA; 33871 MW; ED7027BFE6F86D6F CRC64;
MKITVLGCGA LGQLWLTALC KQGHEVQGWL RVPQPYCSVN LVETDGSIFN ESLTANDPDF
LATSDLLLVT LKAWQVSDAV KSLASTLPVT TPILLIHNGM GTIEELQNIQ QPLLMGTTTH
AARRDGNVII HVANGITHIG PARQQDGDYS YLADILQTVL PDVAWHNNIR AELWRKLAVN
CVINPLTAIW NCPNGELRHH PQEIMQICEE VAAVIEREGH HTSAEDLRDY VMQVIDATAE
NISSMLQDIR ALRHTEIDYI NGFLLRRARA HGIAVPENTR LFEMVKRKES EYERIGTGLP
RPW
