FLK_STRCT
ID FLK_STRCT Reviewed; 139 AA.
AC Q1EMV2;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 11-JUL-2006, sequence version 1.
DT 03-AUG-2022, entry version 46.
DE RecName: Full=Fluoroacetyl-CoA thioesterase;
DE EC=3.1.2.29;
GN Name=flK;
OS Streptomyces cattleya.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=29303;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=16720268; DOI=10.1016/j.chembiol.2006.02.014;
RA Huang F., Haydock S.F., Spiteller D., Mironenko T., Li T.-L., O'Hagan D.,
RA Leadlay P.F., Spencer J.B.;
RT "The gene cluster for fluorometabolite biosynthesis in Streptomyces
RT cattleya: a thioesterase confers resistance to fluoroacetyl-coenzyme A.";
RL Chem. Biol. 13:475-484(2006).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEX WITH COENZYME A AND
RP SUBSTRATE, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF VAL-23; LEU-26;
RP PHE-33; PHE-36; THR-42; GLU-50 AND HIS-76, SUBUNIT, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=20836570; DOI=10.1021/bi101102u;
RA Weeks A.M., Coyle S.M., Jinek M., Doudna J.A., Chang M.C.Y.;
RT "Structural and biochemical studies of a fluoroacetyl-CoA-specific
RT thioesterase reveal a molecular basis for fluorine selectivity.";
RL Biochemistry 49:9269-9279(2010).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) IN COMPLEX WITH COENZYME A AND
RP SUBSTRATE, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF
RP THR-42; GLU-50 AND HIS-76.
RX PubMed=20430898; DOI=10.1074/jbc.m110.107177;
RA Dias M.V.B., Huang F., Chirgadze D.Y., Tosin M., Spiteller D., Dry E.F.V.,
RA Leadlay P.F., Spencer J.B., Blundell T.L.;
RT "Structural basis for the activity and substrate specificity of
RT fluoroacetyl-CoA thioesterase FlK.";
RL J. Biol. Chem. 285:22495-22504(2010).
CC -!- FUNCTION: Hydrolyzes fluoroacetyl-CoA before it can react with citrate
CC synthase, and thus confers fluoroacetate resistance. Cannot use acetyl-
CC CoA as substrate. {ECO:0000269|PubMed:16720268,
CC ECO:0000269|PubMed:20836570}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=fluoroacetyl-CoA + H2O = CoA + fluoroacetate + H(+);
CC Xref=Rhea:RHEA:28542, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:18172, ChEBI:CHEBI:57287, ChEBI:CHEBI:61623; EC=3.1.2.29;
CC Evidence={ECO:0000269|PubMed:16720268, ECO:0000269|PubMed:20836570};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=30 uM for fluoroacetyl-CoA (at pH 8 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:16720268, ECO:0000269|PubMed:20430898};
CC KM=8 uM for fluoroacetyl-CoA (at pH 7.6 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:20836570};
CC KM=2.1 mM for acetyl-CoA (at pH 7.6 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:20836570};
CC Vmax=15 umol/min/mg enzyme with fluoroacetyl-CoA as substrate (at pH
CC 8 and 25 degrees Celsius) {ECO:0000269|PubMed:16720268};
CC Note=kcat is 390 sec(-1) with fluoroacetyl-CoA as substrate and 0.06
CC sec(-1) with acetyl-CoA (at pH 7.6 and 25 degrees Celsius,
CC PubMed:20836570). kcat is 0.044 sec(-1) with fluoroacetyl-CoA as
CC substrate (PubMed:20430898). {ECO:0000269|PubMed:20430898};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:20430898,
CC ECO:0000269|PubMed:20836570}.
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DR EMBL; AM055586; CAJ20012.1; -; Genomic_DNA.
DR PDB; 3KUV; X-ray; 1.50 A; A/B=1-139.
DR PDB; 3KUW; X-ray; 1.90 A; A/B=1-139.
DR PDB; 3KV7; X-ray; 1.56 A; A/B=1-139.
DR PDB; 3KV8; X-ray; 1.85 A; A/B=1-139.
DR PDB; 3KVI; X-ray; 1.76 A; A/B=1-139.
DR PDB; 3KVU; X-ray; 2.00 A; A/B/C/D=1-139.
DR PDB; 3KVZ; X-ray; 2.10 A; A/B/C/D/E/F/G/H=1-139.
DR PDB; 3KW1; X-ray; 1.90 A; A/B/C/D/E/F/G/H=1-139.
DR PDB; 3KX7; X-ray; 1.70 A; A/B=1-139.
DR PDB; 3KX8; X-ray; 2.35 A; A/B/C/D/E/F/G/H=1-139.
DR PDB; 3P2Q; X-ray; 1.85 A; A/B=1-139.
DR PDB; 3P2R; X-ray; 2.46 A; A/B=1-139.
DR PDB; 3P2S; X-ray; 1.95 A; A/B=1-139.
DR PDB; 3P3F; X-ray; 2.30 A; A/B/C/D/E/F=1-139.
DR PDB; 3P3I; X-ray; 2.00 A; A/B/C/D/E/F=1-139.
DR PDBsum; 3KUV; -.
DR PDBsum; 3KUW; -.
DR PDBsum; 3KV7; -.
DR PDBsum; 3KV8; -.
DR PDBsum; 3KVI; -.
DR PDBsum; 3KVU; -.
DR PDBsum; 3KVZ; -.
DR PDBsum; 3KW1; -.
DR PDBsum; 3KX7; -.
DR PDBsum; 3KX8; -.
DR PDBsum; 3P2Q; -.
DR PDBsum; 3P2R; -.
DR PDBsum; 3P2S; -.
DR PDBsum; 3P3F; -.
DR PDBsum; 3P3I; -.
DR AlphaFoldDB; Q1EMV2; -.
DR SMR; Q1EMV2; -.
DR OMA; VFATPAM; -.
DR BioCyc; MetaCyc:MON-15928; -.
DR BRENDA; 3.1.2.29; 5990.
DR EvolutionaryTrace; Q1EMV2; -.
DR GO; GO:0016289; F:CoA hydrolase activity; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR InterPro; IPR025540; FlK.
DR InterPro; IPR029069; HotDog_dom_sf.
DR PANTHER; PTHR36934; PTHR36934; 1.
DR PIRSF; PIRSF014972; FlK; 1.
DR SUPFAM; SSF54637; SSF54637; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase.
FT CHAIN 1..139
FT /note="Fluoroacetyl-CoA thioesterase"
FT /id="PRO_0000419130"
FT ACT_SITE 42
FT ACT_SITE 50
FT ACT_SITE 76
FT BINDING 40..50
FT /ligand="substrate"
FT BINDING 69
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT BINDING 69
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT BINDING 76..77
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT BINDING 120
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT MUTAGEN 23
FT /note="V->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:20836570"
FT MUTAGEN 26
FT /note="L->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:20836570"
FT MUTAGEN 33
FT /note="F->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:20836570"
FT MUTAGEN 36
FT /note="F->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:20836570"
FT MUTAGEN 42
FT /note="T->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT MUTAGEN 42
FT /note="T->C,S: Enhancement of acetyl-CoA binding, but
FT reduced activity toward fluoroacetyl-CoA."
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT MUTAGEN 50
FT /note="E->A,Q: Reduced activity and affinity."
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT MUTAGEN 76
FT /note="H->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:20430898,
FT ECO:0000269|PubMed:20836570"
FT STRAND 10..16
FT /evidence="ECO:0007829|PDB:3KUV"
FT HELIX 19..21
FT /evidence="ECO:0007829|PDB:3KUV"
FT HELIX 23..26
FT /evidence="ECO:0007829|PDB:3KUV"
FT HELIX 31..33
FT /evidence="ECO:0007829|PDB:3KUV"
FT STRAND 38..40
FT /evidence="ECO:0007829|PDB:3KX7"
FT HELIX 42..56
FT /evidence="ECO:0007829|PDB:3KUV"
FT HELIX 58..60
FT /evidence="ECO:0007829|PDB:3KUV"
FT STRAND 65..75
FT /evidence="ECO:0007829|PDB:3KUV"
FT STRAND 84..96
FT /evidence="ECO:0007829|PDB:3KUV"
FT STRAND 99..107
FT /evidence="ECO:0007829|PDB:3KUV"
FT STRAND 109..124
FT /evidence="ECO:0007829|PDB:3KUV"
FT HELIX 125..133
FT /evidence="ECO:0007829|PDB:3KUV"
SQ SEQUENCE 139 AA; 15257 MW; C32DE9465552909C CRC64;
MKDGMRVGER FTHDFVVPPH KTVRHLYPES PEFAEFPEVF ATGFMVGLME WACVRAMAPY
LEPGEGSLGT AICVTHTAAT PPGLTVTVTA ELRSVEGRRL SWRVSAHDGV DEIGSGTHER
AVIHLEKFNA KVRQKTPAG
