HCMA_AQUTE
ID HCMA_AQUTE Reviewed; 562 AA.
AC I3VE77;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 05-SEP-2012, sequence version 1.
DT 03-AUG-2022, entry version 35.
DE RecName: Full=2-hydroxyisobutanoyl-CoA mutase large subunit {ECO:0000305};
DE EC=5.4.99.64 {ECO:0000269|PubMed:22433853, ECO:0000269|PubMed:25720495};
DE AltName: Full=2-hydroxyisobutyryl-CoA mutase large subunit {ECO:0000303|PubMed:22433853};
DE Short=HCM large subunit {ECO:0000303|PubMed:22433853};
GN Name=hcmA {ECO:0000303|PubMed:22433853};
OS Aquincola tertiaricarbonis.
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Aquincola.
OX NCBI_TaxID=391953;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, DISRUPTION PHENOTYPE, AND
RP MUTAGENESIS OF ILE-90.
RC STRAIN=L108;
RX PubMed=22433853; DOI=10.1074/jbc.m111.314690;
RA Yaneva N., Schuster J., Schafer F., Lede V., Przybylski D., Paproth T.,
RA Harms H., Muller R.H., Rohwerder T.;
RT "Bacterial acyl-CoA mutase specifically catalyzes coenzyme B12-dependent
RT isomerization of 2-hydroxyisobutyryl-CoA and (S)-3-hydroxybutyryl-CoA.";
RL J. Biol. Chem. 287:15502-15511(2012).
RN [2] {ECO:0007744|PDB:4R3U}
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) IN COMPLEX WITH HCMB;
RP 2-HYDROXYISOBUTANOYL-COA; 3-HYDROXYBUTANOYL-COA AND COBALAMIN, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND MUTAGENESIS
RP OF ILE-90 AND ASP-117.
RC STRAIN=L108;
RX PubMed=25720495; DOI=10.1074/jbc.m115.645689;
RA Kurteva-Yaneva N., Zahn M., Weichler M.T., Starke R., Harms H.,
RA Muller R.H., Strater N., Rohwerder T.;
RT "Structural basis of the stereospecificity of bacterial B12-dependent 2-
RT hydroxyisobutyryl-CoA mutase.";
RL J. Biol. Chem. 290:9727-9737(2015).
CC -!- FUNCTION: Together with HcmB, catalyzes the isomerization of 2-
CC hydroxyisobutyryl-CoA and 3-hydroxybutyryl-CoA. Is specific for 2-
CC hydroxyisobutyryl-CoA and (S)-3-hydroxybutyryl-CoA, and shows only very
CC low activity with (R)-3-hydroxybutyryl-CoA, isobutyryl-CoA and butyryl-
CC CoA (PubMed:22433853, PubMed:25720495). In vitro, can isomerize
CC pivalyl-CoA and isovaleryl-CoA, with much lower efficiency
CC (PubMed:25720495). Plays a central role in the degradation of
CC substrates bearing a tert-butyl moiety, such as the fuel oxygenate
CC methyl tert-butyl ether (MTBE) and its metabolites (PubMed:22433853).
CC {ECO:0000269|PubMed:22433853, ECO:0000269|PubMed:25720495}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-hydroxyisobutanoyl-CoA = (3S)-3-hydroxybutanoyl-CoA;
CC Xref=Rhea:RHEA:49592, ChEBI:CHEBI:57316, ChEBI:CHEBI:131780;
CC EC=5.4.99.64; Evidence={ECO:0000269|PubMed:22433853,
CC ECO:0000269|PubMed:25720495};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49593;
CC Evidence={ECO:0000269|PubMed:22433853};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=128 uM for (S)-3-hydroxybutyryl-CoA {ECO:0000269|PubMed:22433853};
CC KM=104 uM for 2-hydroxyisobutyryl-CoA {ECO:0000269|PubMed:22433853};
CC KM=1660 uM for (R)-3-hydroxybutyryl-CoA
CC {ECO:0000269|PubMed:22433853};
CC KM=3340 uM for butyryl-CoA {ECO:0000269|PubMed:22433853};
CC KM=550 uM for isobutyryl-CoA {ECO:0000269|PubMed:22433853};
CC KM=374 uM for pivalyl-CoA {ECO:0000269|PubMed:25720495};
CC KM=288 uM for isovaleryl-CoA {ECO:0000269|PubMed:25720495};
CC Vmax=140 nmol/min/mg enzyme with (S)-3-hydroxybutyryl-CoA as
CC substrate {ECO:0000269|PubMed:22433853};
CC Vmax=36 nmol/min/mg enzyme with 2-hydroxyisobutyryl-CoA as substrate
CC {ECO:0000269|PubMed:22433853};
CC Vmax=2.4 nmol/min/mg enzyme with (R)-3-hydroxybutyryl-CoA as
CC substrate {ECO:0000269|PubMed:22433853};
CC Vmax=2.4 nmol/min/mg enzyme with butyryl-CoA as substrate
CC {ECO:0000269|PubMed:22433853};
CC Vmax=0.67 nmol/min/mg enzyme with isobutyryl-CoA as substrate
CC {ECO:0000269|PubMed:22433853};
CC Vmax=11.8 nmol/min/mg enzyme with pivalyl-CoA as substrate
CC {ECO:0000269|PubMed:25720495};
CC Vmax=8.2 nmol/min/mg enzyme with isovaleryl-CoA as substrate
CC {ECO:0000269|PubMed:25720495};
CC Note=kcat is 12 min(-1) with (S)-3-hydroxybutyryl-CoA as substrate.
CC kcat is 3.0 min(-1) with 2-hydroxyisobutyryl-CoA as substrate. kcat
CC is 0.20 min(-1) with (R)-3-hydroxybutyryl-CoA as substrate. kcat is
CC 0.20 min(-1) with butyryl-CoA as substrate. kcat is 0.06 min(-1) with
CC isobutyryl-CoA as substrate (PubMed:22433853). kcat is 0.98 min(-1)
CC with pivalyl-CoA as substrate. kcat is 0.68 min(-1) with isovaleryl-
CC CoA as substrate (PubMed:25720495). {ECO:0000269|PubMed:22433853,
CC ECO:0000269|PubMed:25720495};
CC pH dependence:
CC Optimum pH is 6.6. {ECO:0000269|PubMed:22433853};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius.
CC {ECO:0000269|PubMed:22433853};
CC -!- SUBUNIT: Homotetramer composed of two large substrate-binding subunits
CC (HcmA) and two small cobalamin-binding subunits (HcmB).
CC {ECO:0000269|PubMed:22433853, ECO:0000269|PubMed:25720495}.
CC -!- DISRUPTION PHENOTYPE: Insertion mutant cannot grow on 2-
CC hydroxyisobutyric acid (2-HIBA). {ECO:0000269|PubMed:22433853}.
CC -!- SIMILARITY: Belongs to the acyl-CoA mutase large subunit family.
CC {ECO:0000305}.
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DR EMBL; JQ708092; AFK77668.1; -; Genomic_DNA.
DR PDB; 4R3U; X-ray; 2.50 A; A/B=1-562.
DR PDBsum; 4R3U; -.
DR SMR; I3VE77; -.
DR KEGG; ag:AFK77668; -.
DR BioCyc; MetaCyc:MON-19834; -.
DR BRENDA; 5.4.99.64; 14508.
DR GO; GO:0031419; F:cobalamin binding; IEA:InterPro.
DR GO; GO:0004494; F:methylmalonyl-CoA mutase activity; IEA:InterPro.
DR InterPro; IPR016176; Cbl-dep_enz_cat.
DR InterPro; IPR006099; MeMalonylCoA_mutase_a/b_cat.
DR InterPro; IPR006098; MMCoA_mutase_a_cat.
DR Pfam; PF01642; MM_CoA_mutase; 1.
DR SUPFAM; SSF51703; SSF51703; 1.
DR TIGRFAMs; TIGR00641; acid_CoA_mut_N; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Isomerase.
FT CHAIN 1..562
FT /note="2-hydroxyisobutanoyl-CoA mutase large subunit"
FT /id="PRO_0000455118"
FT BINDING 76..79
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 86..88
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 117
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 196..198
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 235
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 240
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 245
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT BINDING 284
FT /ligand="(3S)-3-hydroxybutanoyl-CoA"
FT /ligand_id="ChEBI:CHEBI:57316"
FT /evidence="ECO:0000269|PubMed:25720495,
FT ECO:0007744|PDB:4R3U"
FT SITE 117
FT /note="Important for the stereospecificity of catalysis"
FT /evidence="ECO:0000305|PubMed:25720495"
FT MUTAGEN 90
FT /note="I->A: 6-fold decrease in catalytic efficiency with
FT 2-hydroxyisobutyryl-CoA as substrate. 320-fold decrease in
FT catalytic efficiency with (S)-3-hydroxybuytryl-CoA as
FT substrate. 6-fold increase in catalytic efficiency with
FT (R)-3-hydroxybutyryl-CoA as substrate. No change in
FT catalytic efficiencies with pivalyl-CoA and isovaleryl-CoA
FT as substrates."
FT /evidence="ECO:0000269|PubMed:25720495"
FT MUTAGEN 90
FT /note="I->F,Y: Loss of activity."
FT /evidence="ECO:0000269|PubMed:22433853"
FT MUTAGEN 90
FT /note="I->L: 37-fold decrease in catalytic efficiency with
FT 2-hydroxyisobutyryl-CoA as substrate. 290-fold decrease in
FT catalytic efficiency with (S)-3-hydroxybuytryl-CoA as
FT substrate. Does not show any significant activities with
FT pivalyl-CoA and isovaleryl-CoA."
FT /evidence="ECO:0000269|PubMed:25720495"
FT MUTAGEN 90
FT /note="I->V: 100-fold decrease in catalytic efficiency with
FT (S)-3-hydroxybuytryl-CoA as substrate. No change in
FT catalytic efficiencies with pivalyl-CoA and isovaleryl-CoA
FT as substrates."
FT /evidence="ECO:0000269|PubMed:22433853"
FT MUTAGEN 117
FT /note="D->A: 2-fold increase in catalytic efficiency with
FT 2-hydroxyisobutyryl-CoA as substrate. Small increase in
FT catalytic efficiency with (S)-3-hydroxybuytryl-CoA as
FT substrate. 1800-fold increase in catalytic efficiency with
FT (R)-3-hydroxybutyryl-CoA as substrate."
FT /evidence="ECO:0000269|PubMed:25720495"
FT MUTAGEN 117
FT /note="D->V: 1.5-fold increase in catalytic efficiency with
FT 2-hydroxyisobutyryl-CoA as substrate. 3-fold decrease in
FT catalytic efficiency with (S)-3-hydroxybuytryl-CoA as
FT substrate. 1300-fold increase in catalytic efficiency with
FT (R)-3-hydroxybutyryl-CoA as substrate. 74-fold increase in
FT catalytic efficiency with pivalyl-CoA as substrate."
FT /evidence="ECO:0000269|PubMed:25720495"
SQ SEQUENCE 562 AA; 63414 MW; 2AA6E3A1314A5C8B CRC64;
MTWLEPQIKS QLQSERKDWE ANEVGAFLKK APERKEQFHT IGDFPVQRTY TAADIADTPL
EDIGLPGRYP FTRGPYPTMY RSRTWTMRQI AGFGTGEDTN KRFKYLIAQG QTGISTDFDM
PTLMGYDSDH PMSDGEVGRE GVAIDTLADM EALLADIDLE KISVSFTINP SAWILLAMYV
ALGEKRGYDL NKLSGTVQAD ILKEYMAQKE YIYPIAPSVR IVRDIITYSA KNLKRYNPIN
ISGYHISEAG SSPLQEAAFT LANLITYVNE VTKTGMHVDE FAPRLAFFFV SQGDFFEEVA
KFRALRRCYA KIMKERFGAR NPESMRLRFH CQTAAATLTK PQYMVNVVRT SLQALSAVLG
GAQSLHTNGY DEAFAIPTED AMKMALRTQQ IIAEESGVAD VIDPLGGSYY VEALTTEYEK
KIFEILEEVE KRGGTIKLIE QGWFQKQIAD FAYETALRKQ SGQKPVIGVN RFVENEEDVK
IEIHPYDNTT AERQISRTRR VRAERDEAKV QAMLDQLVAV AKDESQNLMP LTIELVKAGA
TMGDIVEKLK GIWGTYRETP VF
