ILVC_SLAES
ID ILVC_SLAES Reviewed; 342 AA.
AC D0WGK0;
DT 06-JUL-2016, integrated into UniProtKB/Swiss-Prot.
DT 19-JAN-2010, sequence version 1.
DT 03-AUG-2022, entry version 68.
DE RecName: Full=Ketol-acid reductoisomerase (NADP(+)) {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
DE Short=KARI {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
DE EC=1.1.1.86 {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000269|PubMed:23776225};
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:23776225};
DE AltName: Full=Ketol-acid reductoisomerase type I {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
GN Name=ilvC {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000312|EMBL:EEZ61613.1};
GN ORFNames=HMPREF0762_00954 {ECO:0000312|EMBL:EEZ61613.1};
OS Slackia exigua (strain ATCC 700122 / DSM 15923 / CIP 105133 / JCM 11022 /
OS KCTC 5966 / S-7).
OC Bacteria; Actinobacteria; Coriobacteriia; Eggerthellales; Eggerthellaceae;
OC Slackia.
OX NCBI_TaxID=649764;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700122 / DSM 15923 / CIP 105133 / JCM 11022 / KCTC 5966 / S-7;
RA Weinstock G., Sodergren E., Clifton S., Fulton L., Fulton B., Courtney L.,
RA Fronick C., Harrison M., Strong C., Farmer C., Delahaunty K., Markovic C.,
RA Hall O., Minx P., Tomlinson C., Mitreva M., Nelson J., Hou S., Wollam A.,
RA Pepin K.H., Johnson M., Bhonagiri V., Nash W.E., Warren W., Chinwalla A.,
RA Mardis E.R., Wilson R.K.;
RL Submitted (OCT-2009) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.39 ANGSTROMS) OF WILD-TYPE AND MUTANT IN COMPLEX
RP WITH NADH; NADPH; MAGNESIUM IONS AND SUBSTRATE ANALOG, FUNCTION, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF SER-61 AND SER-63,
RP COFACTOR, AND SUBUNIT.
RX PubMed=23776225; DOI=10.1073/pnas.1306073110;
RA Brinkmann-Chen S., Flock T., Cahn J.K., Snow C.D., Brustad E.M.,
RA McIntosh J.A., Meinhold P., Zhang L., Arnold F.H.;
RT "General approach to reversing ketol-acid reductoisomerase cofactor
RT dependence from NADPH to NADH.";
RL Proc. Natl. Acad. Sci. U.S.A. 110:10946-10951(2013).
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.
CC {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000269|PubMed:23776225}.
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:23776225};
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:23776225};
CC Note=Binds 2 magnesium ions per subunit. {ECO:0000255|HAMAP-
CC Rule:MF_00435};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1 uM for NADPH (at pH 7) {ECO:0000269|PubMed:23776225};
CC KM=45 uM for NADH (at pH 7) {ECO:0000269|PubMed:23776225};
CC Note=kcat is 0.8 sec(-1) for reductoisomerase activity with NADPH as
CC substrate (at pH 7). kcat is 0.41 sec(-1) for reductoisomerase
CC activity with NADH as substrate (at pH 7).
CC {ECO:0000269|PubMed:23776225};
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:23776225}.
CC -!- SIMILARITY: Belongs to the ketol-acid reductoisomerase family.
CC {ECO:0000255|HAMAP-Rule:MF_00435}.
CC -!- CAUTION: The sequence shown here is derived from an EMBL/GenBank/DDBJ
CC whole genome shotgun (WGS) entry which is preliminary data.
CC {ECO:0000312|EMBL:EEZ61613.1}.
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DR EMBL; ACUX02000006; EEZ61613.1; -; Genomic_DNA.
DR PDB; 4KQW; X-ray; 1.39 A; A/B=1-342.
DR PDB; 4KQX; X-ray; 1.80 A; A/B=1-342.
DR PDBsum; 4KQW; -.
DR PDBsum; 4KQX; -.
DR AlphaFoldDB; D0WGK0; -.
DR SMR; D0WGK0; -.
DR STRING; 649764.HMPREF0762_00954; -.
DR EnsemblBacteria; EEZ61613; EEZ61613; HMPREF0762_00954.
DR eggNOG; COG0059; Bacteria.
DR HOGENOM; CLU_033821_0_1_11; -.
DR BRENDA; 1.1.1.383; 9370.
DR UniPathway; UPA00047; UER00056.
DR UniPathway; UPA00049; UER00060.
DR Proteomes; UP000006001; Unassembled WGS sequence.
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..342
FT /note="Ketol-acid reductoisomerase (NADP(+))"
FT /id="PRO_0000436834"
FT DOMAIN 12..192
FT /note="KARI N-terminal Rossmann"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01197"
FT DOMAIN 193..338
FT /note="KARI C-terminal knotted"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01198"
FT ACT_SITE 118
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 35..38
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT BINDING 58
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT BINDING 61
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT BINDING 63
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT BINDING 93..96
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT BINDING 144
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT BINDING 201
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:23776225"
FT BINDING 201
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 205
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:23776225"
FT BINDING 237
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 241
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 262
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435,
FT ECO:0000269|PubMed:23776225"
FT MUTAGEN 61
FT /note="S->D: Inversion of cofactor specificity from NADPH
FT to NADH. Strong decrease of the affinity for NADPH and 2.5-
FT fold decrease of the affinity for NADH. 8-fold decrease of
FT the catalytic efficiency for NADPH and 2-fold increase of
FT the catalytic efficiency for NADH; when associated with D-
FT 63."
FT /evidence="ECO:0000269|PubMed:23776225"
FT MUTAGEN 63
FT /note="S->D: Inversion of cofactor specificity from NADPH
FT to NADH. Strong decrease of the affinity for NADPH and 2.5-
FT fold decrease of the affinity for NADH. 8-fold decrease of
FT the catalytic efficiency for NADPH and 2-fold increase of
FT the catalytic efficiency for NADH; when associated with D-
FT 61."
FT /evidence="ECO:0000269|PubMed:23776225"
FT HELIX 17..19
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 23..26
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 30..33
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 37..48
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 53..57
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 62..69
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 73..76
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 77..83
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 85..89
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 93..95
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 96..103
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 105..107
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 113..119
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 120..123
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 133..142
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 144..152
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 159..165
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 167..169
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 171..181
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 184..187
FT /evidence="ECO:0007829|PDB:4KQW"
FT STRAND 189..191
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 194..207
FT /evidence="ECO:0007829|PDB:4KQW"
FT TURN 208..210
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 211..226
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 231..238
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 240..252
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 254..260
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 263..276
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 279..293
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 296..305
FT /evidence="ECO:0007829|PDB:4KQW"
FT TURN 306..309
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 310..320
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 323..333
FT /evidence="ECO:0007829|PDB:4KQW"
FT HELIX 336..342
FT /evidence="ECO:0007829|PDB:4KQX"
SQ SEQUENCE 342 AA; 37779 MW; 1B129CDB1291A2F1 CRC64;
MSVKTKEKEM AVTILYEQDV DPKVIQGLKV GIIGYGSQGH AHALNLMDSG VDVRVGLREG
SSSWKTAEEA GLKVTDMDTA AEEADVIMVL VPDEIQPKVY QEHIAAHLKA GNTLAFAHGF
NIHYGYIVPP EDVNVIMCAP KGPGHIVRRQ FTEGSGVPDL ACVQQDATGN AWDIVLSYCW
GVGGARSGII KATFAEETEE DLFGEQAVLC GGLVELVKAG FETLTEAGYP PELAYFECYH
EMKMIVDLMY ESGIHFMNYS ISNTAEYGEY YAGPKVINEQ SREAMKEILK RIQDGSFAQE
FVDDCNNGHK RLLEQREAIN THPIETTGAQ IRSMFSWIKK ED
