HOSA_THET2
ID HOSA_THET2 Reviewed; 376 AA.
AC O87198;
DT 30-MAY-2000, integrated into UniProtKB/Swiss-Prot.
DT 24-MAY-2004, sequence version 2.
DT 03-AUG-2022, entry version 115.
DE RecName: Full=Homocitrate synthase {ECO:0000303|PubMed:19996101};
DE Short=HCS {ECO:0000303|PubMed:19996101};
DE EC=2.3.3.14 {ECO:0000269|PubMed:19996101};
GN Name=lys20 {ECO:0000303|PubMed:9868782}; OrderedLocusNames=TT_C1550;
OS Thermus thermophilus (strain ATCC BAA-163 / DSM 7039 / HB27).
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=262724;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, DISRUPTION PHENOTYPE, AND
RP PATHWAY.
RC STRAIN=ATCC BAA-163 / DSM 7039 / HB27;
RX PubMed=9868782; DOI=10.1111/j.1574-6968.1998.tb13341.x;
RA Kosuge T., Hoshino T.;
RT "Lysine is synthesized through the alpha-aminoadipate pathway in Thermus
RT thermophilus.";
RL FEMS Microbiol. Lett. 169:361-367(1998).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC BAA-163 / DSM 7039 / HB27;
RX PubMed=15064768; DOI=10.1038/nbt956;
RA Henne A., Brueggemann H., Raasch C., Wiezer A., Hartsch T., Liesegang H.,
RA Johann A., Lienard T., Gohl O., Martinez-Arias R., Jacobi C.,
RA Starkuviene V., Schlenczeck S., Dencker S., Huber R., Klenk H.-P.,
RA Kramer W., Merkl R., Gottschalk G., Fritz H.-J.;
RT "The genome sequence of the extreme thermophile Thermus thermophilus.";
RL Nat. Biotechnol. 22:547-553(2004).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, AND SUBUNIT.
RC STRAIN=ATCC BAA-163 / DSM 7039 / HB27;
RX PubMed=12095615; DOI=10.1016/s0014-5793(02)02877-6;
RA Wulandari A.P., Miyazaki J., Kobashi N., Nishiyama M., Hoshino T.,
RA Yamane H.;
RT "Characterization of bacterial homocitrate synthase involved in lysine
RT biosynthesis.";
RL FEBS Lett. 522:35-40(2002).
RN [4] {ECO:0007744|PDB:2ZTJ, ECO:0007744|PDB:2ZTK, ECO:0007744|PDB:2ZYF, ECO:0007744|PDB:3A9I}
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEXES WITH (2R)-HOMOCITRATE;
RP 2-OXOGLUTARATE; COPPER; L-LYSINE AND MAGNESIUM, FUNCTION, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, ACTIVITY REGULATION,
RP MUTAGENESIS OF HIS-72, REACTION MECHANISM, AND ACTIVE SITE.
RX PubMed=19996101; DOI=10.1074/jbc.m109.086330;
RA Okada T., Tomita T., Wulandari A.P., Kuzuyama T., Nishiyama M.;
RT "Mechanism of substrate recognition and insight into feedback inhibition of
RT homocitrate synthase from Thermus thermophilus.";
RL J. Biol. Chem. 285:4195-4205(2010).
CC -!- FUNCTION: Catalyzes the aldol-type condensation of 2-oxoglutarate with
CC acetyl-CoA to yield homocitrate (PubMed:19996101, PubMed:12095615).
CC Carries out the first step of the alpha-aminoadipate (AAA) lysine
CC biosynthesis pathway (PubMed:9868782). To a lesser extent, can also use
CC oxaloacetate in place of 2-oxoglutarate, leading to citrate. Does not
CC display 2-isopropylmalate synthase activity since it cannot use 2-
CC oxoisovalerate (PubMed:12095615). {ECO:0000269|PubMed:12095615,
CC ECO:0000269|PubMed:19996101, ECO:0000269|PubMed:9868782}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + acetyl-CoA + H2O = (2R)-homocitrate + CoA +
CC H(+); Xref=Rhea:RHEA:12929, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:57287, ChEBI:CHEBI:57288,
CC ChEBI:CHEBI:58884; EC=2.3.3.14;
CC Evidence={ECO:0000269|PubMed:19996101};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:12930;
CC Evidence={ECO:0000305|PubMed:19996101};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=acetyl-CoA + H2O + oxaloacetate = citrate + CoA + H(+);
CC Xref=Rhea:RHEA:16845, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16452, ChEBI:CHEBI:16947, ChEBI:CHEBI:57287,
CC ChEBI:CHEBI:57288; Evidence={ECO:0000269|PubMed:12095615};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:19996101};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:19996101};
CC -!- ACTIVITY REGULATION: Is highly and competitively inhibited by lysine
CC that binds to the active site and competes with 2-oxoglutarate
CC (PubMed:12095615, PubMed:19996101). Is also slightly inhibited by
CC arginine and 2-aminoethylcysteine (PubMed:12095615).
CC {ECO:0000269|PubMed:12095615, ECO:0000269|PubMed:19996101}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=44 uM for 2-oxoglutarate {ECO:0000269|PubMed:12095615};
CC KM=32 uM for acetyl-CoA (in the presence of 2-oxoglutarate)
CC {ECO:0000269|PubMed:12095615};
CC KM=255 uM for oxaloacetate (in the presence of KCl, necessary for
CC activity) {ECO:0000269|PubMed:12095615};
CC KM=28 uM for acetyl-CoA (in the presence of oxaloacetate and KCl)
CC {ECO:0000269|PubMed:12095615};
CC KM=5.4 uM for 2-oxoglutarate {ECO:0000269|PubMed:19996101};
CC KM=33 uM for acetyl-CoA {ECO:0000269|PubMed:19996101};
CC Note=kcat is 92 min(-1) using 2-oxoglutarate as substrate
CC (PubMed:12095615). kcat is 58 min(-1) using oxaloacetate as substrate
CC (in the presence of KCl, necessary for activity) (PubMed:12095615).
CC kcat is 41 min(-1) using 2-oxoglutarate as substrate
CC (PubMed:19996101). {ECO:0000269|PubMed:12095615,
CC ECO:0000269|PubMed:19996101};
CC Temperature dependence:
CC Optimum temperature is 60 degrees Celsius. Activity is rapidly lost
CC above 70 degrees Celsius. {ECO:0000269|PubMed:12095615};
CC -!- PATHWAY: Amino-acid biosynthesis; L-lysine biosynthesis via AAA
CC pathway; L-alpha-aminoadipate from 2-oxoglutarate: step 1/5.
CC {ECO:0000269|PubMed:9868782, ECO:0000305|PubMed:19996101}.
CC -!- SUBUNIT: Exists in an equilibrium between monomer and homodimer.
CC {ECO:0000269|PubMed:12095615}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene show lysine auxotrophy,
CC which can be complemented with alpha-aminoadipate but not with
CC diaminopimelate. {ECO:0000269|PubMed:9868782}.
CC -!- SIMILARITY: Belongs to the alpha-IPM synthase/homocitrate synthase
CC family. Homocitrate synthase LYS20/LYS21 subfamily. {ECO:0000255|HAMAP-
CC Rule:MF_02222, ECO:0000305}.
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DR EMBL; AB018379; BAA33785.1; -; Genomic_DNA.
DR EMBL; AE017221; AAS81892.1; -; Genomic_DNA.
DR PIR; T51170; T51170.
DR RefSeq; WP_011173924.1; NC_005835.1.
DR PDB; 2ZTJ; X-ray; 1.80 A; A=1-376.
DR PDB; 2ZTK; X-ray; 1.96 A; A=1-376.
DR PDB; 2ZYF; X-ray; 2.15 A; A=1-376.
DR PDB; 3A9I; X-ray; 1.80 A; A=1-376.
DR PDBsum; 2ZTJ; -.
DR PDBsum; 2ZTK; -.
DR PDBsum; 2ZYF; -.
DR PDBsum; 3A9I; -.
DR AlphaFoldDB; O87198; -.
DR SMR; O87198; -.
DR STRING; 262724.TT_C1550; -.
DR EnsemblBacteria; AAS81892; AAS81892; TT_C1550.
DR GeneID; 3170147; -.
DR KEGG; tth:TT_C1550; -.
DR eggNOG; COG0119; Bacteria.
DR HOGENOM; CLU_022158_2_2_0; -.
DR OMA; DWSNGMR; -.
DR OrthoDB; 840579at2; -.
DR BioCyc; MetaCyc:MON-6722; -.
DR SABIO-RK; O87198; -.
DR UniPathway; UPA00033; UER00028.
DR EvolutionaryTrace; O87198; -.
DR PRO; PR:O87198; -.
DR Proteomes; UP000000592; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0036440; F:citrate synthase activity; IEA:RHEA.
DR GO; GO:0004410; F:homocitrate synthase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0019878; P:lysine biosynthetic process via aminoadipic acid; IEA:UniProtKB-UniRule.
DR Gene3D; 3.20.20.70; -; 1.
DR HAMAP; MF_02222; Homocitr_synth_fung_arch; 1.
DR InterPro; IPR002034; AIPM/Hcit_synth_CS.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR011872; Homocitrate_synth_fun/arc.
DR InterPro; IPR000891; PYR_CT.
DR Pfam; PF00682; HMGL-like; 1.
DR TIGRFAMs; TIGR02146; LysS_fung_arch; 1.
DR PROSITE; PS00815; AIPM_HOMOCIT_SYNTH_1; 1.
DR PROSITE; PS00816; AIPM_HOMOCIT_SYNTH_2; 1.
DR PROSITE; PS50991; PYR_CT; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Amino-acid biosynthesis; Cytoplasm; Lysine biosynthesis;
KW Magnesium; Manganese; Metal-binding; Transferase.
FT CHAIN 1..376
FT /note="Homocitrate synthase"
FT /id="PRO_0000140455"
FT DOMAIN 4..259
FT /note="Pyruvate carboxyltransferase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01151"
FT ACT_SITE 292
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:19996101"
FT BINDING 12
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZTJ, ECO:0007744|PDB:2ZYF"
FT BINDING 13
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZYF"
FT BINDING 72
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZTJ, ECO:0007744|PDB:2ZYF"
FT BINDING 92
FT /ligand="L-lysine"
FT /ligand_id="ChEBI:CHEBI:32551"
FT /ligand_note="inhibitor"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:3A9I"
FT BINDING 133
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZTJ, ECO:0007744|PDB:2ZYF"
FT BINDING 135
FT /ligand="L-lysine"
FT /ligand_id="ChEBI:CHEBI:32551"
FT /ligand_note="inhibitor"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:3A9I"
FT BINDING 166
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZTJ, ECO:0007744|PDB:2ZYF"
FT BINDING 166
FT /ligand="L-lysine"
FT /ligand_id="ChEBI:CHEBI:32551"
FT /ligand_note="inhibitor"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:3A9I"
FT BINDING 195
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZYF"
FT BINDING 197
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:19996101,
FT ECO:0007744|PDB:2ZYF"
FT MUTAGEN 72
FT /note="H->L: Significant decrease in sensitivity to lysine
FT inhibition. Large decrease in affinity for 2-oxoglutarate.
FT Almost no effect on affinity for acetyl-CoA and on turnover
FT number."
FT /evidence="ECO:0000269|PubMed:19996101"
FT CONFLICT 104
FT /note="A -> P (in Ref. 1; BAA33785)"
FT /evidence="ECO:0000305"
FT STRAND 5..11
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 13..16
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 24..37
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 40..44
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 47..49
FT /evidence="ECO:0007829|PDB:2ZYF"
FT HELIX 51..61
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 66..75
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 77..85
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 89..95
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 109..126
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 130..136
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT TURN 137..141
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 144..154
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 155..157
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 159..165
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 172..186
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT TURN 187..189
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 190..197
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 203..212
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT STRAND 217..221
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 222..224
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 234..244
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 246..252
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 255..257
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 258..269
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT TURN 278..280
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT TURN 282..285
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 290..298
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 300..303
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 308..311
FT /evidence="ECO:0007829|PDB:2ZTJ"
FT HELIX 342..357
FT /evidence="ECO:0007829|PDB:3A9I"
FT HELIX 363..374
FT /evidence="ECO:0007829|PDB:3A9I"
SQ SEQUENCE 376 AA; 42159 MW; 1B8EEA63C82FCF06 CRC64;
MREWKIIDST LREGEQFEKA NFSTQDKVEI AKALDEFGIE YIEVTTPVAS PQSRKDAEVL
ASLGLKAKVV THIQCRLDAA KVAVETGVQG IDLLFGTSKY LRAAHGRDIP RIIEEAKEVI
AYIREAAPHV EVRFSAEDTF RSEEQDLLAV YEAVAPYVDR VGLADTVGVA TPRQVYALVR
EVRRVVGPRV DIEFHGHNDT GCAIANAYEA IEAGATHVDT TILGIGERNG ITPLGGFLAR
MYTLQPEYVR RKYKLEMLPE LDRMVARMVG VEIPFNNYIT GETAFSHKAG MHLKAIYINP
EAYEPYPPEV FGVKRKLIIA SRLTGRHAIK ARAEELGLHY GEEELHRVTQ HIKALADRGQ
LTLEELDRIL REWITA
