ID AK_THETH Reviewed; 405 AA.
AC P61489; P77991; P97151;
DT 24-MAY-2004, integrated into UniProtKB/Swiss-Prot.
DT 24-MAY-2004, sequence version 1.
DT 27-MAR-2024, entry version 93.
DE RecName: Full=Aspartokinase;
DE EC=2.7.2.4;
DE AltName: Full=Aspartate kinase;
DE Short=AK;
DE Short=ASK;
DE AltName: Full=Threonine-sensitive AK;
DE Short=ThrA;
GN Name=ask; Synonyms=askAB;
OS Thermus thermophilus.
OC Bacteria; Deinococcota; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=274;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, AND NOMENCLATURE.
RC STRAIN=ATCC 33923 / DSM 674 / AT-62;
RX PubMed=7773416; DOI=10.1099/13500872-141-5-1211;
RA Nishiyama M., Kukimoto M., Beppu T., Horiouchi S.;
RT "An operon encoding aspartokinase and purine phosphoribosyltransferase in
RT Thermus flavus.";
RL Microbiology 141:1211-1219(1995).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF LYS-7; GLY-9; GLY-10; SER-41;
RP ALA-42; THR-47; GLU-74; GLY-135; ARG-150; ASP-154; ASP-174 AND ASP-182,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY REGULATION.
RX PubMed=16232547; DOI=10.1016/s1389-1723(99)80146-7;
RA Kobashi N., Nishiyama M., Tanokura M.;
RT "Kinetic and mutation analyses of aspartate kinase from Thermus flavus.";
RL J. Biosci. Bioeng. 87:739-745(1999).
RN [3]
RP ACTIVITY REGULATION, AND SUBUNIT.
RX PubMed=15033471; DOI=10.1016/j.bbrc.2004.02.122;
RA Kato C., Kurihara T., Kobashi N., Yamane H., Nishiyama M.;
RT "Conversion of feedback regulation in aspartate kinase by domain
RT exchange.";
RL Biochem. Biophys. Res. Commun. 316:802-808(2004).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) OF 245-405 IN COMPLEX WITH SUBSTRATE
RP ANALOGS, AND SUBUNIT.
RX PubMed=19490113; DOI=10.1111/j.1742-4658.2009.07030.x;
RA Yoshida A., Tomita T., Kono H., Fushinobu S., Kuzuyama T., Nishiyama M.;
RT "Crystal structures of the regulatory subunit of Thr-sensitive aspartate
RT kinase from Thermus thermophilus.";
RL FEBS J. 276:3124-3136(2009).
CC -!- FUNCTION: Catalyzes the phosphorylation of the beta-carboxyl group of
CC aspartic acid with ATP to yield 4-phospho-L-aspartate, which is
CC involved in the branched biosynthetic pathway leading to the
CC biosynthesis of amino acids threonine, isoleucine and methionine.
CC {ECO:0000269|PubMed:16232547, ECO:0000269|PubMed:7773416}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-aspartate = 4-phospho-L-aspartate + ADP;
CC Xref=Rhea:RHEA:23776, ChEBI:CHEBI:29991, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:57535, ChEBI:CHEBI:456216; EC=2.7.2.4;
CC Evidence={ECO:0000269|PubMed:16232547, ECO:0000269|PubMed:7773416};
CC -!- ACTIVITY REGULATION: Inhibited by threonine.
CC {ECO:0000269|PubMed:15033471, ECO:0000269|PubMed:16232547,
CC ECO:0000269|PubMed:7773416}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.27 mM for aspartate {ECO:0000269|PubMed:16232547};
CC KM=0.5 mM for ATP {ECO:0000269|PubMed:16232547};
CC Note=kcat is 1500 min(-1).;
CC Temperature dependence:
CC Thermostable. {ECO:0000269|PubMed:16232547};
CC -!- PATHWAY: Amino-acid biosynthesis; L-lysine biosynthesis via DAP
CC pathway; (S)-tetrahydrodipicolinate from L-aspartate: step 1/4.
CC -!- PATHWAY: Amino-acid biosynthesis; L-methionine biosynthesis via de novo
CC pathway; L-homoserine from L-aspartate: step 1/3.
CC -!- PATHWAY: Amino-acid biosynthesis; L-threonine biosynthesis; L-threonine
CC from L-aspartate: step 1/5.
CC -!- SUBUNIT: Heterotetramer consisting of 2 isoforms Alpha (catalytic and
CC regulation) and of a homodimer of 2 isoforms Beta (regulation and
CC thermostability). {ECO:0000269|PubMed:15033471,
CC ECO:0000269|PubMed:19490113}.
CC -!- ALTERNATIVE PRODUCTS:
CC Event=Alternative initiation; Named isoforms=2;
CC Name=Alpha; Synonyms=Aspartokinase subunit alpha;
CC IsoId=P61489-1; Sequence=Displayed;
CC Name=Beta; Synonyms=Aspartokinase subunit beta;
CC IsoId=P61489-2; Sequence=VSP_018665;
CC -!- SIMILARITY: Belongs to the aspartokinase family. {ECO:0000305}.
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DR EMBL; D37928; BAA07146.1; -; Genomic_DNA.
DR EMBL; D37928; BAA07147.1; -; Genomic_DNA.
DR RefSeq; WP_024118988.1; NZ_AP024270.1.
DR PDB; 2DT9; X-ray; 2.15 A; A/B=245-405.
DR PDB; 2ZHO; X-ray; 2.98 A; A/B/C/D/E/F=245-405.
DR PDBsum; 2DT9; -.
DR PDBsum; 2ZHO; -.
DR AlphaFoldDB; P61489; -.
DR SMR; P61489; -.
DR BRENDA; 2.7.2.4; 2305.
DR SABIO-RK; P61489; -.
DR UniPathway; UPA00034; UER00015.
DR UniPathway; UPA00050; UER00461.
DR UniPathway; UPA00051; UER00462.
DR EvolutionaryTrace; P61489; -.
DR GO; GO:0004072; F:aspartate kinase activity; IEA:UniProtKB-EC.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0019877; P:diaminopimelate biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0009089; P:lysine biosynthetic process via diaminopimelate; IEA:UniProtKB-UniPathway.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
DR GO; GO:0009088; P:threonine biosynthetic process; IEA:UniProtKB-UniPathway.
DR CDD; cd04261; AAK_AKii-LysC-BS; 1.
DR CDD; cd04923; ACT_AK-LysC-DapG-like_2; 1.
DR CDD; cd04913; ACT_AKii-LysC-BS-like_1; 1.
DR Gene3D; 3.30.70.260; -; 2.
DR Gene3D; 3.40.1160.10; Acetylglutamate kinase-like; 1.
DR InterPro; IPR036393; AceGlu_kinase-like_sf.
DR InterPro; IPR045865; ACT-like_dom_sf.
DR InterPro; IPR002912; ACT_dom.
DR InterPro; IPR041740; AKii-LysC-BS.
DR InterPro; IPR001048; Asp/Glu/Uridylate_kinase.
DR InterPro; IPR005260; Asp_kin_monofn.
DR InterPro; IPR001341; Asp_kinase.
DR InterPro; IPR018042; Aspartate_kinase_CS.
DR InterPro; IPR027795; CASTOR_ACT_dom.
DR NCBIfam; TIGR00656; asp_kin_monofn; 1.
DR NCBIfam; TIGR00657; asp_kinases; 1.
DR PANTHER; PTHR21499; ASPARTATE KINASE; 1.
DR PANTHER; PTHR21499:SF3; ASPARTOKINASE; 1.
DR Pfam; PF00696; AA_kinase; 1.
DR Pfam; PF01842; ACT; 1.
DR Pfam; PF13840; ACT_7; 1.
DR PIRSF; PIRSF000726; Asp_kin; 1.
DR SUPFAM; SSF55021; ACT-like; 2.
DR SUPFAM; SSF53633; Carbamate kinase-like; 1.
DR PROSITE; PS51671; ACT; 1.
DR PROSITE; PS00324; ASPARTOKINASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alternative initiation; Amino-acid biosynthesis; ATP-binding;
KW Diaminopimelate biosynthesis; Kinase; Lysine biosynthesis;
KW Nucleotide-binding; Repeat; Transferase.
FT CHAIN 1..405
FT /note="Aspartokinase"
FT /id="PRO_0000002389"
FT DOMAIN 263..342
FT /note="ACT 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01007"
FT DOMAIN 344..405
FT /note="ACT 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01007"
FT BINDING 7..10
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 25..30
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 41
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 47..49
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 74
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 125..126
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 150..153
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 153
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 173..174
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 179..184
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 209
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250"
FT BINDING 270
FT /ligand="substrate"
FT BINDING 274..275
FT /ligand="substrate"
FT BINDING 288..290
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 294
FT /ligand="substrate"
FT BINDING 355..356
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 369..370
FT /ligand="substrate"
FT BINDING 376..377
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT SITE 7
FT /note="Contribution to the catalysis"
FT SITE 74
FT /note="Contribution to the catalysis"
FT VAR_SEQ 1..244
FT /note="Missing (in isoform Beta)"
FT /evidence="ECO:0000305"
FT /id="VSP_018665"
FT MUTAGEN 7
FT /note="K->A: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 7
FT /note="K->M: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 9
FT /note="G->M: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 10
FT /note="G->A: Significant decrease in the catalytic
FT efficiency."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 41
FT /note="S->A: Significant decrease in the catalytic
FT efficiency. Requires higher concentration of magnesium ion
FT than wild-type."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 42
FT /note="A->S: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 47
FT /note="T->A: Significant decrease in the affinity for
FT aspartic acid. Requires higher concentration of magnesium
FT ion than wild-type."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 74
FT /note="E->A: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 74
FT /note="E->Q: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 135
FT /note="G->A: Very low catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 135
FT /note="G->S: Loss of aspartokinase activity."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 150
FT /note="R->A: Significant decrease in the catalytic
FT efficiency."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 154
FT /note="D->A: Significant decrease in the catalytic
FT efficiency. Requires higher concentration of magnesium ion
FT than wild-type."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 154
FT /note="D->N: Significant decrease in the catalytic
FT efficiency. Requires higher concentration of magnesium ion
FT than wild-type."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 174
FT /note="D->A: Significant decrease in the catalytic
FT efficiency."
FT /evidence="ECO:0000269|PubMed:16232547"
FT MUTAGEN 182
FT /note="D->A: Significant decrease in the catalytic
FT efficiency.Requires higher concentration of magnesium ion
FT than wild-type."
FT /evidence="ECO:0000269|PubMed:16232547"
FT STRAND 251..257
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 259..269
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 274..285
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 292..294
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 303..311
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 312..314
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 315..329
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 332..336
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 338..348
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 349..351
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 354..365
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 372..375
FT /evidence="ECO:0007829|PDB:2DT9"
FT STRAND 377..385
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 386..388
FT /evidence="ECO:0007829|PDB:2DT9"
FT HELIX 389..399
FT /evidence="ECO:0007829|PDB:2DT9"
SQ SEQUENCE 405 AA; 43319 MW; 47A3A29E2B82D0EA CRC64;
MALVVQKYGG TSVGDLERIH KVAQRIAHYR EKGHRLAVVV SAMGHTTDEL IALAKRVNPR
PPFRELDLLT TTGEQVSVAL LSMQLWAMGI PAKGFVQHQI GITTDGRYGD ARILEVNPAR
IREALDQGFV AVIAGFMGTT PEGEITTLGR GGSDTTAVAI AAALGAKECE IYTDTEGVYT
TDPHLIPEAR KLSVIGYDQM LEMAALGARV LHPRAVYYAK RYGVVLHVRS SFSYNPGTLV
KEVAMEMDKA VTGVALDLDH AQIGLIGIPD QPGIAAKVFQ ALAERGIAVD MIIQGVPGHD
PSRQQMAFTV KKDFAQEALE ALEPVLAEIG GEAILRPDIA KVSIVGVGLA STPEVPAKMF
QAVASTGANI EMIATSEVRI SVIIPAEYAE AALRAVHQAF ELDKA
//
