ILVC_ECOLI
ID ILVC_ECOLI Reviewed; 491 AA.
AC P05793; Q2M883;
DT 01-NOV-1988, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 4.
DT 03-AUG-2022, entry version 192.
DE RecName: Full=Ketol-acid reductoisomerase (NADP(+)) {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:2653423};
DE Short=KARI {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:2653423};
DE EC=1.1.1.86 {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000269|PubMed:15654896, ECO:0000269|PubMed:21515217, ECO:0000269|PubMed:2653423, ECO:0000269|PubMed:9015391};
DE AltName: Full=Acetohydroxy-acid isomeroreductase {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:2653423};
DE Short=AHIR {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:2653423};
DE AltName: Full=Alpha-keto-beta-hydroxylacyl reductoisomerase {ECO:0000255|HAMAP-Rule:MF_00435};
DE AltName: Full=Ketol-acid reductoisomerase type 2 {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:16322583};
DE AltName: Full=Ketol-acid reductoisomerase type II {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:16322583};
GN Name=ilvC {ECO:0000255|HAMAP-Rule:MF_00435};
GN OrderedLocusNames=b3774, JW3747;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND INDUCTION.
RC STRAIN=K12;
RX PubMed=3003115; DOI=10.1016/s0021-9258(17)35955-0;
RA Wek R.C., Hatfield G.W.;
RT "Nucleotide sequence and in vivo expression of the ilvY and ilvC genes in
RT Escherichia coli K12. Transcription from divergent overlapping promoters.";
RL J. Biol. Chem. 261:2441-2450(1986).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=1379743; DOI=10.1126/science.1379743;
RA Daniels D.L., Plunkett G. III, Burland V.D., Blattner F.R.;
RT "Analysis of the Escherichia coli genome: DNA sequence of the region from
RT 84.5 to 86.5 minutes.";
RL Science 257:771-778(1992).
RN [3]
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 [4]
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 [5]
RP PROTEIN SEQUENCE OF 2-13.
RC STRAIN=K12 / EMG2;
RX PubMed=9298646; DOI=10.1002/elps.1150180807;
RA Link A.J., Robison K., Church G.M.;
RT "Comparing the predicted and observed properties of proteins encoded in the
RT genome of Escherichia coli K-12.";
RL Electrophoresis 18:1259-1313(1997).
RN [6]
RP INDUCTION.
RX PubMed=3062177; DOI=10.1016/0022-2836(88)90199-4;
RA Wek R.C., Hatfield G.W.;
RT "Transcriptional activation at adjacent operators in the divergent-
RT overlapping ilvY and ilvC promoters of Escherichia coli.";
RL J. Mol. Biol. 203:643-663(1988).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP SUBCELLULAR LOCATION, AND PATHWAY.
RX PubMed=2653423; DOI=10.1021/bi00428a012;
RA Chunduru S.K., Mrachko G.T., Calvo K.C.;
RT "Mechanism of ketol acid reductoisomerase--steady-state analysis and metal
RT ion requirement.";
RL Biochemistry 28:486-493(1989).
RN [8]
RP ACTIVITY REGULATION.
RX PubMed=2189496; DOI=10.1021/bi00463a027;
RA Aulabaugh A., Schloss J.V.;
RT "Oxalyl hydroxamates as reaction-intermediate analogues for ketol-acid
RT reductoisomerase.";
RL Biochemistry 29:2824-2830(1990).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF ARG-68; LYS-69; LYS-75 AND ARG-76.
RX PubMed=9015391; DOI=10.1006/abbi.1996.9802;
RA Rane M.J., Calvo K.C.;
RT "Reversal of the nucleotide specificity of ketol acid reductoisomerase by
RT site-directed mutagenesis identifies the NADPH binding site.";
RL Arch. Biochem. Biophys. 338:83-89(1997).
RN [10]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP MUTAGENESIS OF HIS-132; LYS-155; GLU-213; ASP-217; GLU-221; GLU-389;
RP GLU-393 AND SER-414, AND SUBSTRATE SPECIFICITY.
RX PubMed=15654896; DOI=10.1111/j.1742-4658.2004.04506.x;
RA Tyagi R., Lee Y.T., Guddat L.W., Duggleby R.G.;
RT "Probing the mechanism of the bifunctional enzyme ketol-acid
RT reductoisomerase by site-directed mutagenesis of the active site.";
RL FEBS J. 272:593-602(2005).
RN [11]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF ALA-71; ARG-76; SER-78 AND GLN-110.
RX PubMed=21515217; DOI=10.1016/j.ymben.2011.02.004;
RA Bastian S., Liu X., Meyerowitz J.T., Snow C.D., Chen M.M., Arnold F.H.;
RT "Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable
RT anaerobic 2-methylpropan-1-ol production at theoretical yield in
RT Escherichia coli.";
RL Metab. Eng. 13:345-352(2011).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.60 ANGSTROMS), AND SUBUNIT.
RX PubMed=16322583; DOI=10.1110/ps.051791305;
RA Tyagi R., Duquerroy S., Navaza J., Guddat L.W., Duggleby R.G.;
RT "The crystal structure of a bacterial class II ketol-acid reductoisomerase:
RT domain conservation and evolution.";
RL Protein Sci. 14:3089-3100(2005).
RN [13]
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH MAGNESIUM AND NADP,
RP COFACTOR, AND SUBUNIT.
RX PubMed=23036858; DOI=10.1016/j.jmb.2012.09.018;
RA Wong S.H., Lonhienne T.G., Winzor D.J., Schenk G., Guddat L.W.;
RT "Bacterial and plant ketol-acid reductoisomerases have different mechanisms
RT of induced fit during the catalytic cycle.";
RL J. Mol. Biol. 424:168-179(2012).
CC -!- FUNCTION: Involved in the biosynthesis of branched-chain amino acids
CC (BCAA). Catalyzes an alkyl-migration followed by a ketol-acid reduction
CC of (S)-2-acetolactate (S2AL) to yield (R)-2,3-dihydroxy-isovalerate. In
CC the isomerase reaction, S2AL is rearranged via a Mg-dependent methyl
CC migration to produce 3-hydroxy-3-methyl-2-ketobutyrate (HMKB). In the
CC reductase reaction, this 2-ketoacid undergoes a metal-dependent
CC reduction by NADPH to yield (R)-2,3-dihydroxy-isovalerate. Also able to
CC use 2-ketopantoate, 2-ketoisovalerate, 2-ketovalerate, 2-ketobutyrate,
CC 3-hydroxypyruvate, 3-hydroxy-2-ketobutyrate and pyruvate
CC (PubMed:15654896). {ECO:0000269|PubMed:15654896,
CC ECO:0000269|PubMed:21515217, ECO:0000269|PubMed:2653423,
CC ECO:0000269|PubMed:9015391}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R)-2,3-dihydroxy-3-methylbutanoate + NADP(+) = (2S)-2-
CC acetolactate + H(+) + NADPH; Xref=Rhea:RHEA:22068, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:49072, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:58476; EC=1.1.1.86; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_00435, ECO:0000269|PubMed:15654896,
CC ECO:0000269|PubMed:21515217, ECO:0000269|PubMed:2653423,
CC ECO:0000269|PubMed:9015391};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R,3R)-2,3-dihydroxy-3-methylpentanoate + NADP(+) = (S)-2-
CC ethyl-2-hydroxy-3-oxobutanoate + H(+) + NADPH; Xref=Rhea:RHEA:13493,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:49256, ChEBI:CHEBI:49258,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.1.1.86;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00435};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00435,
CC ECO:0000269|PubMed:15654896, ECO:0000269|PubMed:23036858,
CC ECO:0000269|PubMed:2653423};
CC Note=Binds 2 magnesium ions per subunit. {ECO:0000255|HAMAP-
CC Rule:MF_00435, ECO:0000269|PubMed:23036858,
CC ECO:0000269|PubMed:2653423};
CC -!- ACTIVITY REGULATION: Inhibited by N-hydroxy-N-isopropyloxamate (IpOHA).
CC {ECO:0000269|PubMed:2189496}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.04 mM for NADPH {ECO:0000269|PubMed:21515217};
CC KM=0.042 mM for NADP {ECO:0000269|PubMed:9015391};
CC KM=0.073 mM for NADPH {ECO:0000269|PubMed:9015391};
CC KM=0.17 mM for 2-ketopantoate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:15654896};
CC KM=0.206 mM for NADH {ECO:0000269|PubMed:9015391};
CC KM=0.21 mM for 3-hydroxy-2-ketobutyrate (at pH 8 and 37 degrees
CC Celsius) {ECO:0000269|PubMed:15654896};
CC KM=0.25 mM for 2-acetolactate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:15654896};
CC KM=0.27 mM for 3-hydroxy-3-methyl-2-ketobutyrate (at pH 8 and 37
CC degrees Celsius) {ECO:0000269|PubMed:15654896};
CC KM=0.42 mM for magnesium (with S2AL and NADPH as substrates)
CC {ECO:0000269|PubMed:2653423};
CC KM=1.08 mM for NADH {ECO:0000269|PubMed:21515217};
CC KM=1.54 mM for pyruvate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:2653423};
CC KM=2.96 mM for 3-hydroxypyruvate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:2653423};
CC KM=3.15 mM for 2-ketovalerate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:2653423};
CC KM=4.56 mM for 2-ketobutyrate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:2653423};
CC KM=6.91 mM for 2-ketoisovalerate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:2653423};
CC Vmax=5.421 umol/min/mg enzyme with 3-hydroxypyruvate as substrate (at
CC pH 8 and 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Vmax=3.541 umol/min/mg enzyme with 3-hydroxy-3-methyl-2-ketobutyrate
CC as substrate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:15654896};
CC Vmax=2.25 umol/min/mg enzyme with 2-acetolactate as substrate (at pH
CC 8 and 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Vmax=0.599 umol/min/mg enzyme with 3-hydroxy-2-ketobutyrate as
CC substrate (at pH 8 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:15654896};
CC Vmax=0.196 umol/min/mg enzyme with 2-ketopantoate as substrate (at pH
CC 8 and 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Vmax=0.184 umol/min/mg enzyme with 2-ketoisovalerate as substrate (at
CC pH 8 and 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Vmax=0.168 umol/min/mg enzyme with 2-ketobutyrate as substrate (at pH
CC 8 and 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Vmax=0.05 umol/min/mg enzyme with 2-ketovalerate as substrate (at pH
CC 8 and 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Vmax=0.021 umol/min/mg enzyme with pyruvate as substrate (at pH 8 and
CC 37 degrees Celsius) {ECO:0000269|PubMed:15654896};
CC Note=kcat is 7.2 min(-1) for reductoisomerase activity with NADPH as
CC substrate (PubMed:9015391). kcat is 3.1 min(-1) for reductoisomerase
CC activity with NADPH as substrate (PubMed:9015391). kcat is 0.11 min(-
CC 1) for reductoisomerase activity with NADH as substrate
CC (PubMed:9015391). kcat is 5.376 sec(-1) for reductoisomerase activity
CC with 3-hydroxypyruvate as substrate (at pH 8 and 37 degrees Celsius)
CC (PubMed:15654896). kcat is 3.6 sec(-1) for reductoisomerase activity
CC with NADPH as substrate (PubMed:21515217). kcat is 3.511 sec(-1) for
CC reductoisomerase activity with 3-hydroxy-3-methyl-2-ketobutyrate as
CC substrate (at pH 8 and 37 degrees Celsius) (PubMed:15654896). kcat is
CC 2.231 sec(-1) for reductoisomerase activity with 2-acetolactate as
CC substrate (at pH 8 and 37 degrees Celsius) (PubMed:15654896). kcat is
CC 0.594 sec(-1) for reductoisomerase activity with 3-hydroxy-2-
CC ketobutyrate as substrate (at pH 8 and 37 degrees Celsius)
CC (PubMed:15654896). kcat is 0.3 sec(-1) for reductoisomerase activity
CC with NADH as substrate (PubMed:21515217). kcat is 0.194 sec(-1) for
CC reductoisomerase activity with 2-ketopantoate as substrate (at pH 8
CC and 37 degrees Celsius) (PubMed:15654896). kcat is 0.182 sec(-1) for
CC reductoisomerase activity with 2-ketoisovalerate as substrate (at pH
CC 8 and 37 degrees Celsius) (PubMed:15654896). kcat is 0.167 sec(-1)
CC for reductoisomerase activity with 2-ketobutyrate as substrate (at pH
CC 8 and 37 degrees Celsius) (PubMed:15654896). kcat is 0.05 sec(-1) for
CC reductoisomerase activity with 2-ketovalerate as substrate (at pH 8
CC and 37 degrees Celsius) (PubMed:15654896). kcat is 0.021 sec(-1) for
CC reductoisomerase activity with pyruvate as substrate (at pH 8 and 37
CC degrees Celsius) (PubMed:15654896). {ECO:0000269|PubMed:15654896,
CC ECO:0000269|PubMed:21515217, ECO:0000269|PubMed:9015391};
CC -!- PATHWAY: Amino-acid biosynthesis; L-isoleucine biosynthesis; L-
CC isoleucine from 2-oxobutanoate: step 2/4. {ECO:0000255|HAMAP-
CC Rule:MF_00435, ECO:0000305|PubMed:2653423}.
CC -!- PATHWAY: Amino-acid biosynthesis; L-valine biosynthesis; L-valine from
CC pyruvate: step 2/4. {ECO:0000255|HAMAP-Rule:MF_00435,
CC ECO:0000305|PubMed:2653423}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:16322583,
CC ECO:0000269|PubMed:23036858}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:2653423}.
CC -!- INDUCTION: In the presence of acetohydroxybutyrate and acetolactate and
CC by the activator IlvY. {ECO:0000269|PubMed:3003115,
CC ECO:0000269|PubMed:3062177}.
CC -!- SIMILARITY: Belongs to the ketol-acid reductoisomerase family.
CC {ECO:0000255|HAMAP-Rule:MF_00435}.
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DR EMBL; M11689; AAA24029.1; -; Genomic_DNA.
DR EMBL; M87049; AAA67577.1; -; Genomic_DNA.
DR EMBL; U00096; AAC76779.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE77523.1; -; Genomic_DNA.
DR PIR; A65181; ISECKR.
DR RefSeq; NP_418222.1; NC_000913.3.
DR RefSeq; WP_000024939.1; NZ_SSZK01000025.1.
DR PDB; 1YRL; X-ray; 2.60 A; A/B/C/D=1-491.
DR PDB; 3ULK; X-ray; 2.30 A; A/B=1-491.
DR PDBsum; 1YRL; -.
DR PDBsum; 3ULK; -.
DR AlphaFoldDB; P05793; -.
DR SMR; P05793; -.
DR BioGRID; 4263331; 21.
DR BioGRID; 852586; 1.
DR IntAct; P05793; 4.
DR STRING; 511145.b3774; -.
DR BindingDB; P05793; -.
DR ChEMBL; CHEMBL2366462; -.
DR SWISS-2DPAGE; P05793; -.
DR jPOST; P05793; -.
DR PaxDb; P05793; -.
DR PRIDE; P05793; -.
DR EnsemblBacteria; AAC76779; AAC76779; b3774.
DR EnsemblBacteria; BAE77523; BAE77523; BAE77523.
DR GeneID; 948286; -.
DR KEGG; ecj:JW3747; -.
DR KEGG; eco:b3774; -.
DR PATRIC; fig|511145.12.peg.3891; -.
DR EchoBASE; EB0490; -.
DR eggNOG; COG0059; Bacteria.
DR HOGENOM; CLU_551905_0_0_6; -.
DR InParanoid; P05793; -.
DR OMA; CGLLQTG; -.
DR PhylomeDB; P05793; -.
DR BioCyc; EcoCyc:KETOLREDUCTOISOM-MON; -.
DR BioCyc; MetaCyc:KETOLREDUCTOISOM-MON; -.
DR BRENDA; 1.1.1.383; 2026.
DR BRENDA; 1.1.1.86; 2026.
DR SABIO-RK; P05793; -.
DR UniPathway; UPA00047; UER00056.
DR UniPathway; UPA00049; UER00060.
DR EvolutionaryTrace; P05793; -.
DR PRO; PR:P05793; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0032991; C:protein-containing complex; IDA:EcoCyc.
DR GO; GO:0008677; F:2-dehydropantoate 2-reductase activity; IDA:EcoCyc.
DR GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR GO; GO:0004455; F:ketol-acid reductoisomerase activity; IDA:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0050661; F:NADP binding; IDA:UniProtKB.
DR GO; GO:0009097; P:isoleucine biosynthetic process; IMP:EcoCyc.
DR GO; GO:0015940; P:pantothenate biosynthetic process; IMP:EcoCyc.
DR GO; GO:0009099; P:valine biosynthetic process; IMP:EcoCyc.
DR Gene3D; 1.10.1040.10; -; 1.
DR HAMAP; MF_00435; IlvC; 1.
DR InterPro; IPR008927; 6-PGluconate_DH-like_C_sf.
DR InterPro; IPR013328; 6PGD_dom2.
DR InterPro; IPR013023; KARI.
DR InterPro; IPR000506; KARI_C.
DR InterPro; IPR013116; KARI_N.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR PANTHER; PTHR21371; PTHR21371; 2.
DR Pfam; PF01450; IlvC; 2.
DR Pfam; PF07991; IlvN; 1.
DR SUPFAM; SSF48179; SSF48179; 2.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR00465; ilvC; 1.
DR PROSITE; PS51851; KARI_C; 2.
DR PROSITE; PS51850; KARI_N; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Amino-acid biosynthesis;
KW Branched-chain amino acid biosynthesis; Cytoplasm;
KW Direct protein sequencing; Magnesium; Metal-binding; NADP; Oxidoreductase;
KW Reference proteome; Repeat.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:9298646"
FT CHAIN 2..491
FT /note="Ketol-acid reductoisomerase (NADP(+))"
FT /id="PRO_0000151309"
FT DOMAIN 15..208
FT /note="KARI N-terminal Rossmann"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01197,
FT ECO:0000305|PubMed:16322583"
FT DOMAIN 209..344
FT /note="KARI C-terminal knotted 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01198,
FT ECO:0000305|PubMed:16322583"
FT DOMAIN 345..484
FT /note="KARI C-terminal knotted 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01198,
FT ECO:0000305|PubMed:16322583"
FT ACT_SITE 132
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 45..48
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 68
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000305|PubMed:9015391"
FT BINDING 76
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858, ECO:0000305|PubMed:9015391"
FT BINDING 78
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 108..110
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 158
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 217
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 217
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 221
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 389
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 393
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23036858"
FT BINDING 414
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT MUTAGEN 68
FT /note="R->D: Inversion of cofactor specificity from NADPH
FT to NADH; when associated with L-69, V-75 and D-76."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 68
FT /note="R->Q: 18-fold decrease of the catalytic efficiency
FT and 3-fold decrease of the affinity for NADPH."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 69
FT /note="K->L: Does not significantly alter the affinity for
FT NADPH. Slight increase of the catalytic efficiency.
FT Inversion of cofactor specificity from NADPH to NADH; when
FT associated with D-68, V-75 and D-76."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 71
FT /note="A->S: 7- and 2.5-fold increase of the
FT reductoisomerase activity with NADH and NADPH,
FT respectively."
FT /evidence="ECO:0000269|PubMed:21515217"
FT MUTAGEN 75
FT /note="K->Q: 13-fold decrease of the catalytic efficiency
FT and 3-fold increase of the affinity for NADPH."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 75
FT /note="K->V: Inversion of cofactor specificity from NADPH
FT to NADH; when associated with D-68, L-69 and D-76."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 76
FT /note="R->D: 3-fold increase of the reductoisomerase
FT activity with NADH and slight decrease of the
FT reductoisomerase activity with NADPH."
FT /evidence="ECO:0000269|PubMed:21515217"
FT MUTAGEN 76
FT /note="R->D: Strong increase of catalytic efficiency and
FT 2.5-fold increase of the affinity for NADH. 4-fold decrease
FT of the catalytic efficiency and strong decrease of the
FT affinity for NADPH. Inversion of cofactor specificity from
FT NADPH to NADH; when associated with D-68, L-69 and V-75."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 76
FT /note="R->Q: 20-fold decrease of the catalytic efficiency
FT and 5-fold decrease of the affinity for NADPH."
FT /evidence="ECO:0000269|PubMed:9015391"
FT MUTAGEN 78
FT /note="S->D: 12-fold increase of the reductoisomerase
FT activity with NADH and slight decrease of the
FT reductoisomerase activity with NADPH."
FT /evidence="ECO:0000269|PubMed:21515217"
FT MUTAGEN 110
FT /note="Q->V,A: 12- and 2-fold increase of the
FT reductoisomerase activity with NADH and NADPH,
FT respectively."
FT /evidence="ECO:0000269|PubMed:21515217"
FT MUTAGEN 132
FT /note="H->K: Loss of reductoisomerase activity."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 132
FT /note="H->Q: Loss of reductoisomerase activity. The
FT reductase activity with 3-hydroxypyruvate and HMKB is
FT nearly normal, and the isomerase activity decreases 24-
FT fold."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 155
FT /note="K->E,Q: Loss of reductoisomerase activity."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 155
FT /note="K->R: Loss of reductoisomerase activity. The
FT reductase activity with 3-hydroxypyruvate and HMKB is
FT nearly normal, and the isomerase activity decreases 40-
FT fold."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 213
FT /note="E->D: Loss of reductoisomerase activity. 1.5-fold
FT decrease of the reductase activity with 3-hydroxypyruvate
FT and the isomerase activity decreases 48-fold."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 217
FT /note="D->E,N: Loss of reductoisomerase activity."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 221
FT /note="E->D,Q: Loss of reductoisomerase activity."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 389
FT /note="E->D: Loss of reductoisomerase activity. 1.5-fold
FT decrease of the reductase activity with 3-hydroxypyruvate
FT and the isomerase activity decreases 4-fold."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 389
FT /note="E->Q: Loss of reductoisomerase activity."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 393
FT /note="E->D: Loss of reductoisomerase activity. The
FT reductase activity with HMKB is nearly normal."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 414
FT /note="S->A: Loss of reductoisomerase activity. The
FT isomerase activity decreases 15-fold."
FT /evidence="ECO:0000269|PubMed:15654896"
FT MUTAGEN 414
FT /note="S->T: Loss of reductoisomerase activity. The
FT isomerase activity decreases 24-fold."
FT /evidence="ECO:0000269|PubMed:15654896"
FT CONFLICT 251
FT /note="E -> K (in Ref. 1; AAA24029)"
FT /evidence="ECO:0000305"
FT HELIX 5..7
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 10..17
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 20..22
FT /evidence="ECO:0007829|PDB:1YRL"
FT HELIX 25..28
FT /evidence="ECO:0007829|PDB:3ULK"
FT TURN 29..32
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 33..35
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 38..44
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 47..58
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 62..67
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 69..73
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 77..84
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 88..91
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 92..95
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 96..98
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 100..104
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 108..110
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 111..118
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 119..121
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 127..132
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 134..137
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 147..156
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 158..166
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 173..177
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 179..181
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 187..197
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 200..202
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 205..207
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 210..222
FT /evidence="ECO:0007829|PDB:3ULK"
FT TURN 223..226
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 227..242
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 247..275
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 279..309
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 311..321
FT /evidence="ECO:0007829|PDB:3ULK"
FT TURN 322..324
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 325..336
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 338..341
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 351..356
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 359..377
FT /evidence="ECO:0007829|PDB:3ULK"
FT TURN 378..380
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 383..388
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 391..393
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 394..412
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 415..431
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 433..437
FT /evidence="ECO:0007829|PDB:3ULK"
FT STRAND 443..446
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 455..466
FT /evidence="ECO:0007829|PDB:3ULK"
FT HELIX 469..487
FT /evidence="ECO:0007829|PDB:3ULK"
SQ SEQUENCE 491 AA; 54069 MW; 9CA34BA61C9AEBBA CRC64;
MANYFNTLNL RQQLAQLGKC RFMGRDEFAD GASYLQGKKV VIVGCGAQGL NQGLNMRDSG
LDISYALRKE AIAEKRASWR KATENGFKVG TYEELIPQAD LVINLTPDKQ HSDVVRTVQP
LMKDGAALGY SHGFNIVEVG EQIRKDITVV MVAPKCPGTE VREEYKRGFG VPTLIAVHPE
NDPKGEGMAI AKAWAAATGG HRAGVLESSF VAEVKSDLMG EQTILCGMLQ AGSLLCFDKL
VEEGTDPAYA EKLIQFGWET ITEALKQGGI TLMMDRLSNP AKLRAYALSE QLKEIMAPLF
QKHMDDIISG EFSSGMMADW ANDDKKLLTW REETGKTAFE TAPQYEGKIG EQEYFDKGVL
MIAMVKAGVE LAFETMVDSG IIEESAYYES LHELPLIANT IARKRLYEMN VVISDTAEYG
NYLFSYACVP LLKPFMAELQ PGDLGKAIPE GAVDNGQLRD VNEAIRSHAI EQVGKKLRGY
MTDMKRIAVA G
