ILVC_IGNAA
ID ILVC_IGNAA Reviewed; 335 AA.
AC E0SRA9;
DT 06-JUL-2016, integrated into UniProtKB/Swiss-Prot.
DT 02-NOV-2010, sequence version 1.
DT 03-AUG-2022, entry version 68.
DE RecName: Full=Ketol-acid reductoisomerase (NAD(P)(+)) {ECO:0000303|PubMed:25172159};
DE Short=KARI {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:25172159};
DE EC=1.1.1.383 {ECO:0000269|PubMed:25172159, ECO:0000269|PubMed:26644020};
DE AltName: Full=Acetohydroxy-acid isomeroreductase {ECO:0000255|HAMAP-Rule:MF_00435};
DE Short=AHIR {ECO:0000255|HAMAP-Rule:MF_00435};
DE AltName: Full=Alpha-keto-beta-hydroxylacyl reductoisomerase {ECO:0000255|HAMAP-Rule:MF_00435};
DE AltName: Full=Ketol-acid reductoisomerase type 1 {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:25172159};
DE AltName: Full=Ketol-acid reductoisomerase type I {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:25172159};
GN Name=ilvC {ECO:0000255|HAMAP-Rule:MF_00435};
GN OrderedLocusNames=Igag_1561 {ECO:0000312|EMBL:ADM28363.1};
OS Ignisphaera aggregans (strain DSM 17230 / JCM 13409 / AQ1.S1).
OC Archaea; Crenarchaeota; Thermoprotei; Desulfurococcales;
OC Desulfurococcaceae; Ignisphaera.
OX NCBI_TaxID=583356;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 17230 / JCM 13409 / AQ1.S1;
RX PubMed=21304693; DOI=10.4056/sigs.1072907;
RA Goker M., Held B., Lapidus A., Nolan M., Spring S., Yasawong M., Lucas S.,
RA Glavina Del Rio T., Tice H., Cheng J.F., Goodwin L., Tapia R., Pitluck S.,
RA Liolios K., Ivanova N., Mavromatis K., Mikhailova N., Pati A., Chen A.,
RA Palaniappan K., Brambilla E., Land M., Hauser L., Chang Y.J.,
RA Jeffries C.D., Brettin T., Detter J.C., Han C., Rohde M., Sikorski J.,
RA Woyke T., Bristow J., Eisen J.A., Markowitz V., Hugenholtz P.,
RA Kyrpides N.C., Klenk H.P.;
RT "Complete genome sequence of Ignisphaera aggregans type strain (AQ1.S1).";
RL Stand. Genomic Sci. 3:66-75(2010).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBSTRATE
RP SPECIFICITY.
RX PubMed=25172159; DOI=10.1016/j.ymben.2014.08.003;
RA Brinkmann-Chen S., Cahn J.K., Arnold F.H.;
RT "Uncovering rare NADH-preferring ketol-acid reductoisomerases.";
RL Metab. Eng. 26:17-22(2014).
RN [3] {ECO:0007744|PDB:4XDZ, ECO:0007744|PDB:4XEH}
RP X-RAY CRYSTALLOGRAPHY (1.15 ANGSTROMS) IN COMPLEX WITH NADP AND MAGNESIUM
RP IONS, COFACTOR, AND SUBUNIT.
RX PubMed=25849365; DOI=10.1042/bj20150183;
RA Cahn J.K., Brinkmann-Chen S., Spatzal T., Wiig J.A., Buller A.R.,
RA Einsle O., Hu Y., Ribbe M.W., Arnold F.H.;
RT "Cofactor specificity motifs and the induced fit mechanism in class I
RT ketol-acid reductoisomerases.";
RL Biochem. J. 468:475-484(2015).
RN [4] {ECO:0007744|PDB:5E4R}
RP X-RAY CRYSTALLOGRAPHY (1.94 ANGSTROMS) IN COMPLEX WITH NADP, FUNCTION,
RP CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=26644020; DOI=10.1002/pro.2852;
RA Cahn J.K., Brinkmann-Chen S., Buller A.R., Arnold F.H.;
RT "Artificial domain duplication replicates evolutionary history of ketol-
RT acid reductoisomerases.";
RL Protein Sci. 25:1241-1248(2016).
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 or NADH to yield (R)-2,3-dihydroxy-isovalerate.
CC {ECO:0000269|PubMed:25172159, ECO:0000269|PubMed:26644020}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R)-2,3-dihydroxy-3-methylbutanoate + NAD(+) = (2S)-2-
CC acetolactate + H(+) + NADH; Xref=Rhea:RHEA:30627, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:49072, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945,
CC ChEBI:CHEBI:58476; EC=1.1.1.383;
CC Evidence={ECO:0000269|PubMed:25172159, ECO:0000269|PubMed:26644020};
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=1.1.1.383;
CC Evidence={ECO:0000269|PubMed:25172159, ECO:0000269|PubMed:26644020};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00435,
CC ECO:0000269|PubMed:25849365};
CC Note=Binds 2 magnesium ions per subunit. {ECO:0000255|HAMAP-
CC Rule:MF_00435};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.1 mM for S2AL (at pH 7 and 85 degrees Celsius)
CC {ECO:0000269|PubMed:26644020};
CC Note=kcat is 3.3 sec(-1) for reductoisomerase activity with NADPH (at
CC pH 7 and 85 degrees Celsius with S2AL as substrate)
CC (PubMed:26644020). kcat is 0.03 sec(-1) for reductoisomerase activity
CC with NADPH (at pH 7 with S2AL as substrate) (PubMed:25172159). kcat
CC is 0.02 sec(-1) for reductoisomerase activity with NADH (at pH 7 with
CC S2AL as substrate) (PubMed:25172159). {ECO:0000269|PubMed:25172159,
CC ECO:0000269|PubMed:26644020};
CC -!- PATHWAY: Amino-acid biosynthesis; L-isoleucine biosynthesis; L-
CC isoleucine from 2-oxobutanoate: step 2/4. {ECO:0000255|HAMAP-
CC Rule:MF_00435}.
CC -!- PATHWAY: Amino-acid biosynthesis; L-valine biosynthesis; L-valine from
CC pyruvate: step 2/4. {ECO:0000255|HAMAP-Rule:MF_00435}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:25849365}.
CC -!- SIMILARITY: Belongs to the ketol-acid reductoisomerase family.
CC {ECO:0000255|HAMAP-Rule:MF_00435}.
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DR EMBL; CP002098; ADM28363.1; -; Genomic_DNA.
DR PDB; 4XDZ; X-ray; 1.15 A; A/B=1-335.
DR PDB; 4XEH; X-ray; 1.39 A; A=1-335.
DR PDB; 5E4R; X-ray; 1.94 A; A=1-335.
DR PDBsum; 4XDZ; -.
DR PDBsum; 4XEH; -.
DR PDBsum; 5E4R; -.
DR AlphaFoldDB; E0SRA9; -.
DR SMR; E0SRA9; -.
DR STRING; 583356.Igag_1561; -.
DR EnsemblBacteria; ADM28363; ADM28363; Igag_1561.
DR KEGG; iag:Igag_1561; -.
DR HOGENOM; CLU_033821_0_1_2; -.
DR OMA; RAMFSWL; -.
DR BRENDA; 1.1.1.382; 14023.
DR BRENDA; 1.1.1.86; 14023.
DR SABIO-RK; E0SRA9; -.
DR UniPathway; UPA00047; UER00056.
DR UniPathway; UPA00049; UER00060.
DR Proteomes; UP000001304; Chromosome.
DR GO; GO:0004455; F:ketol-acid reductoisomerase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0000287; F:magnesium ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0050661; F:NADP binding; IEA:InterPro.
DR GO; GO:0009097; P:isoleucine biosynthetic process; IEA:UniProtKB-UniRule.
DR GO; GO:0009099; P:valine biosynthetic process; IEA:UniProtKB-UniRule.
DR HAMAP; MF_00435; IlvC; 1.
DR InterPro; IPR008927; 6-PGluconate_DH-like_C_sf.
DR InterPro; IPR013023; KARI.
DR InterPro; IPR000506; KARI_C.
DR InterPro; IPR013116; KARI_N.
DR InterPro; IPR014359; KARI_prok.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR PANTHER; PTHR21371; PTHR21371; 1.
DR Pfam; PF01450; IlvC; 1.
DR Pfam; PF07991; IlvN; 1.
DR PIRSF; PIRSF000116; IlvC_gammaproteo; 1.
DR SUPFAM; SSF48179; SSF48179; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR00465; ilvC; 1.
DR PROSITE; PS51851; KARI_C; 1.
DR PROSITE; PS51850; KARI_N; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Amino-acid biosynthesis;
KW Branched-chain amino acid biosynthesis; Magnesium; Metal-binding; NADP;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..335
FT /note="Ketol-acid reductoisomerase (NAD(P)(+))"
FT /id="PRO_0000436833"
FT DOMAIN 2..182
FT /note="KARI N-terminal Rossmann"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01197"
FT DOMAIN 183..328
FT /note="KARI C-terminal knotted"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01198"
FT ACT_SITE 108
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 25..28
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:25849365, ECO:0007744|PDB:4XDZ,
FT ECO:0007744|PDB:5E4R"
FT BINDING 49
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:25849365,
FT ECO:0007744|PDB:4XDZ, ECO:0007744|PDB:5E4R"
FT BINDING 53
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:25849365, ECO:0007744|PDB:4XDZ,
FT ECO:0007744|PDB:5E4R"
FT BINDING 83..86
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:25849365, ECO:0007744|PDB:4XDZ,
FT ECO:0007744|PDB:5E4R"
FT BINDING 134
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:25849365, ECO:0007744|PDB:4XDZ,
FT ECO:0007744|PDB:5E4R"
FT BINDING 191
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:25849365"
FT BINDING 191
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 195
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:25849365"
FT BINDING 227
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 231
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 252
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT HELIX 7..9
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 13..15
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 19..23
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 27..38
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 42..51
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 52..59
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 67..72
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 75..79
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 83..85
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 86..93
FT /evidence="ECO:0007829|PDB:4XDZ"
FT TURN 94..97
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 103..109
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 110..113
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 125..132
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 134..142
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 149..155
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 157..159
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 161..171
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 174..176
FT /evidence="ECO:0007829|PDB:4XDZ"
FT STRAND 179..181
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 184..197
FT /evidence="ECO:0007829|PDB:4XDZ"
FT TURN 198..200
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 201..216
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 221..228
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 230..250
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 253..266
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 269..283
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 286..296
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 300..310
FT /evidence="ECO:0007829|PDB:4XDZ"
FT HELIX 313..325
FT /evidence="ECO:0007829|PDB:4XDZ"
SQ SEQUENCE 335 AA; 37346 MW; A09760D040982C42 CRC64;
MAKIYKDEDI SLEPIKNKTI AILGYGSQGR AWALNLRDSG LNVVVGLERQ GDSWRRAIDD
GFKPMYTKDA VAIADIIVFL VPDMVQKSLW LNSVKDFMKK GADLVFAHGF NIHFKIIEPP
KDSDVYMIAP KSPGPIVRRS YEMGGGVPAL VAVYQNVSGE ALQKALAIAK GIGCARAGVI
ESTFKEETET DLFGEQVILV GGIMELIKAS FETLVEEGYQ PEVAYFETVN ELKLIVDLIY
EKGLTGMLRA VSDTAKYGGI TVGKFIIDKS VRDKMKIVLE RIRSGEFARE WIKEYERGMP
TVFKELSELE GSTIETVGRK LREMMFRGMK QISSH
