CAMK_RHOSO
ID CAMK_RHOSO Reviewed; 257 AA.
AC Q93TU6;
DT 26-JUN-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 03-AUG-2022, entry version 68.
DE RecName: Full=6-oxocamphor hydrolase;
DE Short=OCH;
DE EC=3.7.1.18;
DE AltName: Full=Beta-diketone hydrolase;
GN Name=camK;
OS Rhodococcus sp.
OC Bacteria; Actinobacteria; Corynebacteriales; Nocardiaceae; Rhodococcus.
OX NCBI_TaxID=1831;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF N-TERMINUS,
RP FUNCTION, CATALYTIC ACTIVITY, MASS SPECTROMETRY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, SUBSTRATE SPECIFICITY, REACTION MECHANISM,
RP AND SUBUNIT.
RC STRAIN=NCIMB 9784;
RX PubMed=11278926; DOI=10.1074/jbc.m011538200;
RA Grogan G., Roberts G.A., Bougioukou D., Turner N.J., Flitsch S.L.;
RT "The desymmetrization of bicyclic beta -diketones by an enzymatic retro-
RT Claisen reaction. A new reaction of the crotonase superfamily.";
RL J. Biol. Chem. 276:12565-12572(2001).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NCIMB 9784;
RX PubMed=15346763; DOI=10.1080/10425170310001656765;
RA Roberts G.A., Grogan G., Turner N.J., Flitsch S.L.;
RT "Nucleotide sequence of a portion of the camphor-degrading gene cluster
RT from Rhodococcus sp. NCIMB 9784.";
RL DNA Seq. 15:96-103(2004).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS), AND SUBUNIT.
RC STRAIN=NCIMB 9784;
RX PubMed=12421807; DOI=10.1074/jbc.m211188200;
RA Whittingham J.L., Turkenburg J.P., Verma C.S., Walsh M.A., Grogan G.;
RT "The 2-A crystal structure of 6-oxo camphor hydrolase. New structural
RT diversity in the crotonase superfamily.";
RL J. Biol. Chem. 278:1744-1750(2003).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF MUTANTS ALA-122 IN COMPLEX WITH
RP SUBSTRATE, MUTAGENESIS OF HIS-122; HIS-145 AND ASP-154, FUNCTION, REACTION
RP MECHANISM, AND SUBUNIT.
RX PubMed=15138275; DOI=10.1074/jbc.m403514200;
RA Leonard P.M., Grogan G.;
RT "Structure of 6-oxo camphor hydrolase H122A mutant bound to its natural
RT product, (2S,4S)-alpha-campholinic acid: mutant structure suggests an
RT atypical mode of transition state binding for a crotonase homolog.";
RL J. Biol. Chem. 279:31312-31317(2004).
CC -!- FUNCTION: Catalyzes the carbon-carbon bond cleavage of the bicyclic
CC beta-diketone 6-oxocamphor via a retro-Claisen reaction to yield the
CC optically active (2R,4S)-beta-campholinic acid. It is also able to
CC cleave 2,2-disubstituted cyclohexa-1,3-diones such as 2-methyl-2-
CC propylcyclohexa-1,3-dione and 2-methyl-2-butylcyclohexa-1,3-dione which
CC result in racemic keto acid products. Transformations of the bicyclic
CC diketone substrates bicyclo[2.2.1]heptane 2,6-dione and
CC bicyclo[2.2.2]octane-2,6-dione yield (S)-keto acid products.
CC {ECO:0000269|PubMed:11278926, ECO:0000269|PubMed:15138275}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=bornane-2,6-dione + H2O = [(1S)-4-hydroxy-2,2,3-
CC trimethylcyclopent-3-enyl]acetate + H(+); Xref=Rhea:RHEA:33415,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:64893,
CC ChEBI:CHEBI:64894; EC=3.7.1.18;
CC Evidence={ECO:0000269|PubMed:11278926};
CC -!- ACTIVITY REGULATION: Inhibited by copper and mercury ions. Also
CC inhibited by thiol active reagents, such as N-ethylmaleimide and
CC hydroxymercuribenzoate. EDTA has a slight activating effect.
CC {ECO:0000269|PubMed:11278926}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.05 mM for 6-oxocamphor {ECO:0000269|PubMed:11278926};
CC Note=kcat is 167 sec(-1) with 6-oxocamphor.;
CC pH dependence:
CC Optimum pH is 7. {ECO:0000269|PubMed:11278926};
CC -!- SUBUNIT: Hexamer. Dimer of trimers. {ECO:0000269|PubMed:11278926,
CC ECO:0000269|PubMed:12421807, ECO:0000269|PubMed:15138275}.
CC -!- MASS SPECTROMETRY: Mass=28488; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:11278926};
CC -!- SIMILARITY: Belongs to the enoyl-CoA hydratase/isomerase family.
CC {ECO:0000305}.
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DR EMBL; AF323755; AAK50622.1; -; Genomic_DNA.
DR PDB; 1O8U; X-ray; 2.00 A; A/B/C/D/E/F=1-257.
DR PDB; 1SZO; X-ray; 1.90 A; A/B/C/D/E/F/G/H/I/J/K/L=1-257.
DR PDBsum; 1O8U; -.
DR PDBsum; 1SZO; -.
DR AlphaFoldDB; Q93TU6; -.
DR SMR; Q93TU6; -.
DR DrugBank; DB02906; (2s,4s)-Alpha-Campholinic Acid.
DR PRIDE; Q93TU6; -.
DR KEGG; ag:AAK50622; -.
DR BioCyc; MetaCyc:MON-17352; -.
DR BRENDA; 3.7.1.18; 5397.
DR SABIO-RK; Q93TU6; -.
DR EvolutionaryTrace; Q93TU6; -.
DR GO; GO:0016823; F:hydrolase activity, acting on acid carbon-carbon bonds, in ketonic substances; IDA:UniProtKB.
DR InterPro; IPR029045; ClpP/crotonase-like_dom_sf.
DR InterPro; IPR001753; Enoyl-CoA_hydra/iso.
DR Pfam; PF00378; ECH_1; 1.
DR SUPFAM; SSF52096; SSF52096; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Hydrolase.
FT CHAIN 1..257
FT /note="6-oxocamphor hydrolase"
FT /id="PRO_0000422737"
FT ACT_SITE 124
FT /note="Nucleophile"
FT /evidence="ECO:0000250"
FT BINDING 40
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15138275"
FT BINDING 145
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15138275"
FT BINDING 154
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15138275"
FT BINDING 244
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15138275"
FT MUTAGEN 45
FT /note="H->A: Great decrease of catalytic efficiency."
FT MUTAGEN 122
FT /note="H->A: Great decrease of catalytic efficiency, and
FT the binding affinity is five times that of the wild-type."
FT /evidence="ECO:0000269|PubMed:15138275"
FT MUTAGEN 145
FT /note="H->A: Very large decrease in the catalytic
FT efficiency. 100-fold decrease in the binding affinity."
FT /evidence="ECO:0000269|PubMed:15138275"
FT MUTAGEN 154
FT /note="D->N: 100-fold decrease in the binding affinity."
FT /evidence="ECO:0000269|PubMed:15138275"
FT HELIX 8..11
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 18..23
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 26..31
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 42..57
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 63..67
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 72..74
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 79..81
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 87..106
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 111..115
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 124..127
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 129..135
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 139..141
FT /evidence="ECO:0007829|PDB:1SZO"
FT TURN 143..145
FT /evidence="ECO:0007829|PDB:1O8U"
FT HELIX 146..148
FT /evidence="ECO:0007829|PDB:1SZO"
FT TURN 153..155
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 156..164
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 166..174
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 181..186
FT /evidence="ECO:0007829|PDB:1SZO"
FT STRAND 191..194
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 196..211
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 215..225
FT /evidence="ECO:0007829|PDB:1SZO"
FT HELIX 227..251
FT /evidence="ECO:0007829|PDB:1SZO"
SQ SEQUENCE 257 AA; 28483 MW; 0AE4A9E54CE83EFF CRC64;
MKQLATPFQE YSQKYENIRL ERDGGVLLVT VHTEGKSLVW TSTAHDELAY CFHDIACDRE
NKVVILTGTG PSFCNEIDFT SFNLGTPHDW DEIIFEGQRL LNNLLSIEVP VIAAVNGPVT
NHPEIPVMSD IVLAAESATF QDGPHFPSGI VPGDGAHVVW PHVLGSNRGR YFLLTGQELD
ARTALDYGAV NEVLSEQELL PRAWELARGI AEKPLLARRY ARKVLTRQLR RVMEADLSLG
LAHEALAAID LGMESEQ
