THIL_CUPNH
ID THIL_CUPNH Reviewed; 393 AA.
AC P14611; Q0KBP8;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-APR-1990, sequence version 1.
DT 03-AUG-2022, entry version 144.
DE RecName: Full=Acetyl-CoA acetyltransferase;
DE EC=2.3.1.9 {ECO:0000269|PubMed:4198758};
DE AltName: Full=Acetoacetyl-CoA thiolase;
DE AltName: Full=Beta-ketothiolase {ECO:0000303|PubMed:2670935};
GN Name=phaA {ECO:0000303|PubMed:1476773};
GN Synonyms=phbA {ECO:0000303|PubMed:2670935}; OrderedLocusNames=H16_A1438;
OS Cupriavidus necator (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442
OS / H16 / Stanier 337) (Ralstonia eutropha).
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Burkholderiaceae; Cupriavidus.
OX NCBI_TaxID=381666;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION, AND FUNCTION.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=2670935; DOI=10.1016/s0021-9258(19)84824-x;
RA Peoples O.P., Sinskey A.J.;
RT "Poly-beta-hydroxybutyrate biosynthesis in Alcaligenes eutrophus H16.
RT Characterization of the genes encoding beta-ketothiolase and acetoacetyl-
RT CoA reductase.";
RL J. Biol. Chem. 264:15293-15297(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=16964242; DOI=10.1038/nbt1244;
RA Pohlmann A., Fricke W.F., Reinecke F., Kusian B., Liesegang H., Cramm R.,
RA Eitinger T., Ewering C., Poetter M., Schwartz E., Strittmatter A., Voss I.,
RA Gottschalk G., Steinbuechel A., Friedrich B., Bowien B.;
RT "Genome sequence of the bioplastic-producing 'Knallgas' bacterium Ralstonia
RT eutropha H16.";
RL Nat. Biotechnol. 24:1257-1262(2006).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-24, FUNCTION IN PHB SYNTHESIS, AND
RP PATHWAY.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=2670936; DOI=10.1016/s0021-9258(19)84825-1;
RA Peoples O.P., Sinskey A.J.;
RT "Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16.
RT Identification and characterization of the PHB polymerase gene (phbC).";
RL J. Biol. Chem. 264:15298-15303(1989).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, SUBUNIT, AND PATHWAY.
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=4198758; DOI=10.1042/bj1340239;
RA Oeding V., Schlegel H.G.;
RT "Beta-ketothiolase from Hydrogenomonas eutropha H16 and its significance in
RT the regulation of poly-beta-hydroxybutyrate metabolism.";
RL Biochem. J. 134:239-248(1973).
RN [5]
RP GENE NAME.
RX PubMed=1476773; DOI=10.1111/j.1574-6968.1992.tb05841.x;
RA Steinbuechel A., Hustede E., Liebergesell M., Pieper U., Timm A.,
RA Valentin H.;
RT "Molecular basis for biosynthesis and accumulation of polyhydroxyalkanoic
RT acids in bacteria.";
RL FEMS Microbiol. Rev. 9:217-230(1992).
RN [6] {ECO:0007744|PDB:4O99, ECO:0007744|PDB:4O9A, ECO:0007744|PDB:4O9C}
RP X-RAY CRYSTALLOGRAPHY (1.52 ANGSTROMS) OF 2-393 APOPROTEIN AND IN COMPLEX
RP WITH COA, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE, SUBUNIT, AND
RP MUTAGENESIS OF CYS-88; HIS-156; PHE-219; SER-248; HIS-349 AND CYS-379.
RX PubMed=25152395; DOI=10.1016/j.bbrc.2014.08.074;
RA Kim E.J., Kim K.J.;
RT "Crystal structure and biochemical characterization of PhaA from Ralstonia
RT eutropha, a polyhydroxyalkanoate-producing bacterium.";
RL Biochem. Biophys. Res. Commun. 452:124-129(2014).
CC -!- FUNCTION: Catalyzes the condensation of two acetyl-coA units to form
CC acetoacetyl-CoA (PubMed:4198758). Is involved in the biosynthesis of
CC polyhydroxybutyrate (PHB), which is accumulated as an intracellular
CC energy reserve material when cells grow under conditions of nutrient
CC limitation (PubMed:2670936, PubMed:4198758). Also catalyzes the reverse
CC reaction, i.e. the cleavage of acetoacetyl-CoA, and is therefore also
CC involved in the reutilization of PHB (PubMed:4198758, PubMed:25152395).
CC {ECO:0000269|PubMed:25152395, ECO:0000269|PubMed:2670935,
CC ECO:0000269|PubMed:2670936, ECO:0000269|PubMed:4198758}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 acetyl-CoA = acetoacetyl-CoA + CoA; Xref=Rhea:RHEA:21036,
CC ChEBI:CHEBI:57286, ChEBI:CHEBI:57287, ChEBI:CHEBI:57288; EC=2.3.1.9;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU10020,
CC ECO:0000269|PubMed:25152395, ECO:0000269|PubMed:4198758};
CC -!- ACTIVITY REGULATION: The condensation reaction is inhibited by free
CC CoA. The cleavage reaction is characterized by substrate inhibition by
CC acetoacetyl-CoA, which is partially relieved by free CoA.
CC {ECO:0000269|PubMed:4198758}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=390 uM for acetyl-CoA {ECO:0000269|PubMed:4198758};
CC pH dependence:
CC Optimum pH is 7.8 for the condensation reaction and 8.1 for the
CC reverse cleavage reaction. {ECO:0000269|PubMed:4198758};
CC -!- PATHWAY: Biopolymer metabolism; poly-(R)-3-hydroxybutanoate
CC biosynthesis. {ECO:0000269|PubMed:2670936, ECO:0000269|PubMed:4198758}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:25152395,
CC ECO:0000269|PubMed:4198758}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- SIMILARITY: Belongs to the thiolase-like superfamily. Thiolase family.
CC {ECO:0000305}.
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DR EMBL; J04987; AAA21972.1; -; Genomic_DNA.
DR EMBL; AM260479; CAJ92573.1; -; Genomic_DNA.
DR EMBL; J05003; AAA21976.1; -; Genomic_DNA.
DR PIR; A34340; XXALAE.
DR RefSeq; WP_010810132.1; NZ_CP039287.1.
DR PDB; 4O99; X-ray; 1.96 A; A/B/C/D=2-393.
DR PDB; 4O9A; X-ray; 1.52 A; A/B/C/D=2-393.
DR PDB; 4O9C; X-ray; 2.00 A; A/B/C/D/E/F/G/H=1-393.
DR PDBsum; 4O99; -.
DR PDBsum; 4O9A; -.
DR PDBsum; 4O9C; -.
DR AlphaFoldDB; P14611; -.
DR SMR; P14611; -.
DR STRING; 381666.H16_A1438; -.
DR EnsemblBacteria; CAJ92573; CAJ92573; H16_A1438.
DR GeneID; 57643537; -.
DR KEGG; reh:H16_A1438; -.
DR eggNOG; COG0183; Bacteria.
DR HOGENOM; CLU_031026_0_0_4; -.
DR OMA; ICPSIAI; -.
DR OrthoDB; 1058688at2; -.
DR BioCyc; MetaCyc:MON-13086; -.
DR BRENDA; 2.3.1.9; 231.
DR UniPathway; UPA00917; -.
DR Proteomes; UP000008210; Chromosome 1.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0003985; F:acetyl-CoA C-acetyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0042619; P:poly-hydroxybutyrate biosynthetic process; IEA:UniProtKB-KW.
DR CDD; cd00751; thiolase; 1.
DR Gene3D; 3.40.47.10; -; 2.
DR InterPro; IPR002155; Thiolase.
DR InterPro; IPR016039; Thiolase-like.
DR InterPro; IPR020615; Thiolase_acyl_enz_int_AS.
DR InterPro; IPR020610; Thiolase_AS.
DR InterPro; IPR020617; Thiolase_C.
DR InterPro; IPR020613; Thiolase_CS.
DR InterPro; IPR020616; Thiolase_N.
DR Pfam; PF02803; Thiolase_C; 1.
DR Pfam; PF00108; Thiolase_N; 1.
DR PIRSF; PIRSF000429; Ac-CoA_Ac_transf; 1.
DR SUPFAM; SSF53901; SSF53901; 2.
DR TIGRFAMs; TIGR01930; AcCoA-C-Actrans; 1.
DR PROSITE; PS00098; THIOLASE_1; 1.
DR PROSITE; PS00737; THIOLASE_2; 1.
DR PROSITE; PS00099; THIOLASE_3; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Cytoplasm; PHB biosynthesis;
KW Reference proteome; Transferase.
FT CHAIN 1..393
FT /note="Acetyl-CoA acetyltransferase"
FT /id="PRO_0000206418"
FT ACT_SITE 88
FT /note="Acyl-thioester intermediate"
FT /evidence="ECO:0000269|PubMed:25152395"
FT ACT_SITE 349
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10020,
FT ECO:0000305|PubMed:25152395"
FT ACT_SITE 379
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10020,
FT ECO:0000305|PubMed:25152395"
FT MUTAGEN 88
FT /note="C->S: Almost complete loss of acetoacetyl-CoA
FT thiolase activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 156
FT /note="H->A: Almost complete loss of acetoacetyl-CoA
FT thiolase activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 219
FT /note="F->A: About 50% loss of acetoacetyl-CoA thiolase
FT activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 219
FT /note="F->Y: 2-fold increase of acetoacetyl-CoA thiolase
FT activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 221
FT /note="R->A: Almost complete loss of acetoacetyl-CoA
FT thiolase activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 248
FT /note="S->A: About 40% loss of acetoacetyl-CoA thiolase
FT activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 349
FT /note="H->A: Almost complete loss of acetoacetyl-CoA
FT thiolase activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT MUTAGEN 379
FT /note="C->S: Almost complete loss of acetoacetyl-CoA
FT thiolase activity."
FT /evidence="ECO:0000269|PubMed:25152395"
FT STRAND 4..12
FT /evidence="ECO:0007829|PDB:4O9A"
FT TURN 20..23
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 26..41
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 45..47
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 50..54
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 65..72
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 81..85
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 87..89
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 90..103
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 108..118
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 123..125
FT /evidence="ECO:0007829|PDB:4O9A"
FT TURN 126..130
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 133..135
FT /evidence="ECO:0007829|PDB:4O9C"
FT STRAND 137..141
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 142..147
FT /evidence="ECO:0007829|PDB:4O9A"
FT TURN 151..154
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 157..168
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 172..191
FT /evidence="ECO:0007829|PDB:4O9A"
FT TURN 192..198
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 202..204
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 207..210
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 212..214
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 226..230
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 244..246
FT /evidence="ECO:0007829|PDB:4O99"
FT STRAND 251..261
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 262..268
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 273..283
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 286..291
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 293..304
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 308..310
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 312..316
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 321..331
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 335..337
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 344..347
FT /evidence="ECO:0007829|PDB:4O9A"
FT TURN 351..353
FT /evidence="ECO:0007829|PDB:4O9A"
FT HELIX 354..369
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 373..380
FT /evidence="ECO:0007829|PDB:4O9A"
FT TURN 381..383
FT /evidence="ECO:0007829|PDB:4O9A"
FT STRAND 384..391
FT /evidence="ECO:0007829|PDB:4O9A"
SQ SEQUENCE 393 AA; 40549 MW; 232C1127E20B3961 CRC64;
MTDVVIVSAA RTAVGKFGGS LAKIPAPELG AVVIKAALER AGVKPEQVSE VIMGQVLTAG
SGQNPARQAA IKAGLPAMVP AMTINKVCGS GLKAVMLAAN AIMAGDAEIV VAGGQENMSA
APHVLPGSRD GFRMGDAKLV DTMIVDGLWD VYNQYHMGIT AENVAKEYGI TREAQDEFAV
GSQNKAEAAQ KAGKFDEEIV PVLIPQRKGD PVAFKTDEFV RQGATLDSMS GLKPAFDKAG
TVTAANASGL NDGAAAVVVM SAAKAKELGL TPLATIKSYA NAGVDPKVMG MGPVPASKRA
LSRAEWTPQD LDLMEINEAF AAQALAVHQQ MGWDTSKVNV NGGAIAIGHP IGASGCRILV
TLLHEMKRRD AKKGLASLCI GGGMGVALAV ERK
