HACL_ACTC0
ID HACL_ACTC0 Reviewed; 590 AA.
AC P0DUV9;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 23-FEB-2022, sequence version 1.
DT 03-AUG-2022, entry version 3.
DE RecName: Full=2-hydroxyacyl-CoA lyase {ECO:0000303|PubMed:34952003};
DE Short=AcHACL {ECO:0000303|PubMed:34952003};
DE Short=HACL {ECO:0000303|PubMed:34952003};
DE EC=4.1.-.- {ECO:0000269|PubMed:34952003};
DE AltName: Full=2-hydroxyisobutyryl-CoA lyase {ECO:0000303|PubMed:32351493};
OS Actinomycetospora chiangmaiensis (strain DSM 45062 / JCM 15998 / NBRC
OS 104400 / YIM 0006).
OC Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae;
OC Actinomycetospora.
OX NCBI_TaxID=1120948;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 45062 / JCM 15998 / NBRC 104400 / YIM 0006;
RA Kyrpides N., Huntemann M., Han J., Chen A., Mavromatis K., Markowitz V.,
RA Palaniappan K., Ivanova N., Schaumberg A., Pati A., Liolios K.,
RA Nordberg H.P., Cantor M.N., Hua S.X., Woyke T.;
RL Submitted (APR-2013) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP IDENTIFICATION BY MASS SPECTROMETRY, FUNCTION, POSSIBLE THIAMINE
RP DIPHOSPHATE COFACTOR, ACTIVITY REGULATION, AND INDUCTION BY 2-HIBA.
RC STRAIN=DSM 45062 / JCM 15998 / NBRC 104400 / YIM 0006;
RX PubMed=32351493; DOI=10.3389/fmicb.2020.00691;
RA Rohwerder T., Rohde M.T., Jehmlich N., Purswani J.;
RT "Actinobacterial Degradation of 2-Hydroxyisobutyric Acid Proceeds via
RT Acetone and Formyl-CoA by Employing a Thiamine-Dependent Lyase Reaction.";
RL Front. Microbiol. 11:691-691(2020).
RN [3]
RP STRUCTURE BY ELECTRON MICROSCOPY (1.55 ANGSTROMS) OF 15-590 IN COMPLEX WITH
RP COFACTORS; SUBSTRATE OR PRODUCT, FUNCTION, CATALYTIC ACTIVITY, PROBABLE
RP REACTION MECHANISM, COFACTORS, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT,
RP DOMAIN, BIOTECHNOLOGY, AND MUTAGENESIS OF GLU-493.
RC STRAIN=DSM 45062 / JCM 15998 / NBRC 104400 / YIM 0006;
RX PubMed=34952003; DOI=10.1016/j.jbc.2021.101522;
RA Zahn M., Koenig G., Pham H.V.C., Seroka B., Lazny R., Yang G.,
RA Ouerfelli O., Lotowski Z., Rohwerder T.;
RT "Mechanistic details of the actinobacterial lyase-catalyzed degradation
RT reaction of 2-hydroxyisobutyryl-CoA.";
RL J. Biol. Chem. 298:101522-101522(2022).
CC -!- FUNCTION: A lyase that reversibly degrades 2-hydroxyisobutyryl-CoA (2-
CC HIB-CoA) to acetone and formyl-CoA (PubMed:34952003). Probably also
CC cleaves 2-hydroxy-2-methylbutyryl-CoA to butanone and formyl-CoA. Does
CC not act on 2-hydroxy-2-ethylbutyryl-CoA (Probable). A C-terminal lid
CC closes the active site upon substrate binding, and with residues Leu-
CC 127 and Ile-492 restricts the size of the active site cavity so it can
CC only use short-chain (C4 and C5) acyl substrates (PubMed:34952003).
CC Part of a pathway that allows cells to grow on 2-methylpropane-1,2-diol
CC or 2-hydroxyisobutyric acid (2-HIBA) as a sole carbon source
CC (PubMed:32351493). {ECO:0000269|PubMed:32351493,
CC ECO:0000269|PubMed:34952003, ECO:0000305|PubMed:32351493}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-hydroxyisobutanoyl-CoA = acetone + formyl-CoA;
CC Xref=Rhea:RHEA:69424, ChEBI:CHEBI:15347, ChEBI:CHEBI:57376,
CC ChEBI:CHEBI:131780; Evidence={ECO:0000269|PubMed:34952003,
CC ECO:0000305|PubMed:32351493};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:69425;
CC Evidence={ECO:0000269|PubMed:34952003};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:34952003};
CC -!- COFACTOR:
CC Name=thiamine diphosphate; Xref=ChEBI:CHEBI:58937;
CC Evidence={ECO:0000269|PubMed:34952003, ECO:0000305|PubMed:32351493};
CC Note=Binds 1 thiamine diphosphate per dimer.
CC {ECO:0000269|PubMed:34952003, ECO:0000305|PubMed:32351493};
CC -!- ACTIVITY REGULATION: Activity is stimulated by thiamine diphosphate.
CC {ECO:0000269|PubMed:32351493}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=120 uM for 2-hydroxyisobutanoyl-CoA {ECO:0000269|PubMed:34952003};
CC Vmax=1200 nmol/min/mg enzyme {ECO:0000269|PubMed:34952003};
CC Note=kcat is 1.3 sec(-1). {ECO:0000269|PubMed:34952003};
CC -!- SUBUNIT: A homotetramer formed by a dimer of dimers; active sites are
CC located in the dimer interface. {ECO:0000269|PubMed:34952003}.
CC -!- INDUCTION: 29-fold induced by growth on 2-hydroxyisobutyric acid (2-
CC HIBA), but not by growth on acetone (at protein level).
CC {ECO:0000269|PubMed:32351493}.
CC -!- DOMAIN: The C-terminal lid closes the active site upon substrate
CC binding. {ECO:0000269|PubMed:34952003}.
CC -!- BIOTECHNOLOGY: This class of enzyme could be engineered to produce
CC biotechnologically interesting condensations of formyl-CoA with
CC aldehydes and ketones. {ECO:0000305|PubMed:34952003}.
CC -!- SIMILARITY: Belongs to the TPP enzyme family. {ECO:0000305}.
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DR EMBL; KB903220; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR RefSeq; WP_018331913.1; NZ_KB903220.1.
DR PDB; 7PT1; X-ray; 1.55 A; A/B=15-588.
DR PDB; 7PT2; X-ray; 1.76 A; A/B=15-588.
DR PDB; 7PT3; X-ray; 1.63 A; A/B=15-588.
DR PDB; 7PT4; X-ray; 1.64 A; A/B=15-590.
DR PDBsum; 7PT1; -.
DR PDBsum; 7PT2; -.
DR PDBsum; 7PT3; -.
DR PDBsum; 7PT4; -.
DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW.
DR GO; GO:0000287; F:magnesium ion binding; IEA:InterPro.
DR GO; GO:0030976; F:thiamine pyrophosphate binding; IEA:InterPro.
DR InterPro; IPR029035; DHS-like_NAD/FAD-binding_dom.
DR InterPro; IPR029061; THDP-binding.
DR InterPro; IPR012000; Thiamin_PyroP_enz_cen_dom.
DR InterPro; IPR012001; Thiamin_PyroP_enz_TPP-bd_dom.
DR InterPro; IPR000399; TPP-bd_CS.
DR InterPro; IPR045229; TPP_enz.
DR InterPro; IPR011766; TPP_enzyme-bd_C.
DR PANTHER; PTHR18968; PTHR18968; 1.
DR Pfam; PF02775; TPP_enzyme_C; 1.
DR Pfam; PF00205; TPP_enzyme_M; 1.
DR Pfam; PF02776; TPP_enzyme_N; 1.
DR SUPFAM; SSF52467; SSF52467; 1.
DR SUPFAM; SSF52518; SSF52518; 2.
DR PROSITE; PS00187; TPP_ENZYMES; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Lyase; Magnesium; Metal-binding; Thiamine pyrophosphate.
FT CHAIN 1..590
FT /note="2-hydroxyacyl-CoA lyase"
FT /id="PRO_0000454895"
FT REGION 566..590
FT /note="C-terminal lid"
FT /evidence="ECO:0000269|PubMed:34952003"
FT ACT_SITE 493
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:34952003"
FT BINDING 43
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 128
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000305|PubMed:34952003"
FT BINDING 255
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 273..274
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 362
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 410..412
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 417
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 433
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 460
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 461..462
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 487..492
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 487
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 489
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:34952003"
FT BINDING 561..564
FT /ligand="2-hydroxyisobutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:131780"
FT /evidence="ECO:0000269|PubMed:34952003"
FT MUTAGEN 493
FT /note="E->A: 10-fold decrease of 2-HIB-CoA cleavage rate,
FT 6-fold increase in KM."
FT /evidence="ECO:0000269|PubMed:34952003"
FT MUTAGEN 493
FT /note="E->K: No cleavage of 2-HIB-CoA."
FT /evidence="ECO:0000269|PubMed:34952003"
FT MUTAGEN 493
FT /note="E->Q: 50-fold decrease of 2-HIB-CoA cleavage rate,
FT 1.5-fold increase in KM."
FT /evidence="ECO:0000269|PubMed:34952003"
SQ SEQUENCE 590 AA; 62765 MW; 12E27DFADDC8D476 CRC64;
MADRQDAERS GAGPARQSVP VASLVAEFLQ EHGVDRVFGL QGGHIQPIWD QLARRGVRIV
DVRDEGSAVH MAHAHTELTG QTAVAMVTAG PGVTNTVTAV ANASVSRIPL LVIGGCPPIP
QSNMGPLQDI PHTAILEPIT RLARTLRSAD QVLREFDEAW ARASGDRGEP GPVYLEIPTD
VLRRDVPPAL QMREHLRAKP KRRPQPHPDD VAAVADLIRA AEKPAIISGR GARTTDGTDL
VRLLDASGAA YLDTQESRGL VPDSHPAAVG SARSAVMRDT DLLITVGRQL DYQLGMGSPA
VFPHAKVVRI ADTASELIDN RRGEVEILAE PGAALAAIAD ALKDHTPDTS WRDELKAKHR
KRAEDYRQAL HSTENGADGH IHPNRIFGAL DALDGDVLDL GETIMIADGG DLLSFGRLGI
TKARRYLDAG AFGCLGVATP FAIGAALAYP DRPVVAVTGD GAFGITATEI DTAVRHDAKI
VVIVSNNRAW NIERYDQAEN YGLVVGTDLA DSDYAGVARA FGAHGERVTD PAELEGAIRR
ALANAPALVD VVTTQDAASP DSGKGLGFVP DYQALTPWND AEVARRQEGI
