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Literature summary extracted from

  • Dias, M.V.; Huang, F.; Chirgadze, D.Y.; Tosin, M.; Spiteller, D.; Dry, E.F.; Leadlay, P.F.; Spencer, J.B.; Blundell, T.L.
    Structural basis for the activity and substrate specificity of fluoroacetyl-CoA thioesterase FlK (2010), J. Biol. Chem., 285, 22495-22504.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.2.29 His-tagged protein expressed in Escherichia coli Rosetta (DE3)pLysS Streptomyces cattleya

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.2.29 FlK forms a dimer, in which each subunit adopts a hot dog fold as observed for type II thioesterases. Unlike other type II thioesterases, FlK employs a catalytic triad composed of Thr42, His76, and a water molecule Streptomyces cattleya
3.1.2.29 sitting drop method, polyethylene glycol monomethyl ether 2000 Streptomyces cattleya

Protein Variants

EC Number Protein Variants Comment Organism
3.1.2.29 E50A disturbed protein folding. 37% of wild-type catalytic efficiency, substrate inhibition above 0.019 mM Streptomyces cattleya
3.1.2.29 E50A putative member of the catalytic triad Streptomyces cattleya
3.1.2.29 H76D insoluble protein Streptomyces cattleya
3.1.2.29 H76D putative member of the catalytic triad, completely insoluble Streptomyces cattleya
3.1.2.29 T42A disturbed protein folding, complete loss of activity Streptomyces cattleya
3.1.2.29 T42A putative member of the catalytic triad Streptomyces cattleya
3.1.2.29 T42S disturbed protein folding. 19fold increase in catalytic efficiency. Substrate inhibition above 0.012 mM Streptomyces cattleya
3.1.2.29 T42S putative member of the catalytic triad Streptomyces cattleya

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.2.29 fluoroacetyl-CoA substrate inhibition of mutants T42S and E50A above 0.012 and 0.019 mM, respectively; T42S mutant protein: substrate inhibition observed above 0.012 mM, E50A mutant protein: substrate inhibition observed above 0.019 mM Streptomyces cattleya

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.2.29 0.015
-
 fluoroacetyl-CoA mutant T42S, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.015
-
 fluoroacetyl-CoA T42S mutant protein, pH not specified in the publication, 25°C Streptomyces cattleya
3.1.2.29 0.03
-
 fluoroacetyl-CoA wild-type, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.03
-
 fluoroacetyl-CoA wild type protein, pH not specified in the publication, 25°C Streptomyces cattleya
3.1.2.29 0.206
-
 fluoroacetyl-CoA mutant E50A, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.206
-
 fluoroacetyl-CoA E50A mutant protein, pH not specified in the publication, 25°C Streptomyces cattleya

Organism

EC Number Organism UniProt Comment Textmining
3.1.2.29 Streptomyces cattleya Q1EMV2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.2.29 immobilized metal ion affinity chromatography (Co2+), His tag removed, gel filtration Streptomyces cattleya

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.2.29 fluoroacetyl-CoA + H2O
-
 Streptomyces cattleya fluoroacetate + CoA
-
 ?
3.1.2.29 fluoroacetyl-CoA + H2O does not accept acetyl-CoA as a substrate Streptomyces cattleya fluoroacetate + CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.2.29 homodimer crystal structure analysis Streptomyces cattleya

Synonyms

EC Number Synonyms Comment Organism
3.1.2.29 fluoroacetyl-CoA thioesterase
-
 Streptomyces cattleya

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.2.29 0.044
-
 fluoroacetyl-CoA wild-type, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.044
-
 fluoroacetyl-CoA wild type protein, pH not specified in the publication, 25°C Streptomyces cattleya
3.1.2.29 0.111
-
 fluoroacetyl-CoA mutant E50A, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.111
-
 fluoroacetyl-CoA E50A mutant protein, pH not specified in the publication, 25°C Streptomyces cattleya
3.1.2.29 0.409
-
 fluoroacetyl-CoA mutant T42S, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.409
-
 fluoroacetyl-CoA T42S mutant protein, pH not specified in the publication, 25°C Streptomyces cattleya

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.2.29 0.54
-
 fluoroacetyl-CoA mutant E50A, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 0.54
-
 fluoroacetyl-CoA E50A mutant protein, pH not specified in the publication, 25°C Streptomyces cattleya
3.1.2.29 1.47
-
 fluoroacetyl-CoA wild-type, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 1.47
-
 fluoroacetyl-CoA wild type protein, pH not specified in the publication, 25°C Streptomyces cattleya
3.1.2.29 27.3
-
 fluoroacetyl-CoA mutant T42S, 25°C, pH not specified in the publication Streptomyces cattleya
3.1.2.29 27.3
-
 fluoroacetyl-CoA T42S mutant protein, pH not specified in the publication, 25°C Streptomyces cattleya