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Literature summary extracted from

  • Weeks, A.M.; Coyle, S.M.; Jinek, M.; Doudna, J.A.; Chang, M.C.
    Structural and biochemical studies of a fluoroacetyl-CoA-specific thioesterase reveal a molecular basis for fluorine selectivity (2010), Biochemistry, 49, 9269-9279.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.2.29 wild-type in complex with fluoroacetate, mutant F36A both alone and as product complex, to 2.3 A, 2.3 A, and 2.0 A resolution, respectively Streptomyces cattleya

Protein Variants

EC Number Protein Variants Comment Organism
3.1.2.29 E50Q about 5000fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 F33A about 1000fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 F36A about 100fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 H76A about 100000fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 L26A about 1000fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 T42A about 10000fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 T42C about 1000fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 T42S about 100fold decrease in catalytic efficiency Streptomyces cattleya
3.1.2.29 V23A about 100fold decrease in catalytic efficiency Streptomyces cattleya

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.2.29 0.001
-
 fluoroacetyl-CoA wild-type, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.014
-
 fluoroacetyl-CoA mutant E50Q, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.02
-
 fluoroacetyl-CoA mutant H76A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.06
-
 fluoroacetyl-CoA mutant T42S, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.061
-
 acetyl-CoA mutant T42S, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.11
-
 fluoroacetyl-CoA mutant T42A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.12
-
 fluoroacetyl-CoA mutant L26A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.3
-
 fluoroacetyl-CoA mutant V23A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.36
-
 fluoroacetyl-CoA mutant T42C, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.39
-
 acetyl-CoA mutant T42C, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.71
-
 fluoroacetyl-CoA mutant F36A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.9
-
 acetyl-CoA mutant V23A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 1.5
-
 fluoroacetyl-CoA mutant F33A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 2.1
-
 acetyl-CoA wild-type, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 3
-
 acetyl-CoA mutant F36A, 25°C, pH 7.6 Streptomyces cattleya

Organism

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.2.29 acetyl-CoA + H2O poor substrate Streptomyces cattleya acetate + CoA
-
 ?
3.1.2.29 fluoroacetyl-CoA + H2O
-
 Streptomyces cattleya fluoroacetate + CoA
-
 ?
3.1.2.29 additional information Thr 42, Glu 50, and His 76 are key catalytic residues. FlK minimizes interaction with the thioester carbonyl, leading to selection against acetyl-CoA binding. Fluorinated substrates bind in a hydrophobic binding pocket created by a lid structure, containing Val 23, Leu 26, Phe 33, and Phe 36, that is not found in other structurally characterized members of this superfamily. Water plays a critical role in fluorine specificity. The Phe 36 gate residue functions to exclude water from the active site Streptomyces cattleya ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.2.29 FlK
-
 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 0.003
-
 fluoroacetyl-CoA mutant H76A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.01
-
 acetyl-CoA mutant T42C, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.01
-
 acetyl-CoA mutant T42S, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.01
-
 acetyl-CoA mutant V23A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.06
-
 acetyl-CoA wild-type, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.1
-
 acetyl-CoA mutant F36A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.13
-
 fluoroacetyl-CoA mutant E50Q, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.45
-
 fluoroacetyl-CoA mutant T42A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 6.7
-
 fluoroacetyl-CoA mutant L26A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 19
-
 fluoroacetyl-CoA mutant T42S, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 27
-
 fluoroacetyl-CoA mutant F33A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 110
-
 fluoroacetyl-CoA mutant T42C, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 150
-
 fluoroacetyl-CoA mutant V23A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 270
-
 fluoroacetyl-CoA mutant F36A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 390
-
 fluoroacetyl-CoA wild-type, 25°C, pH 7.6 Streptomyces cattleya

General Information

EC Number General Information Comment Organism
3.1.2.29 physiological function presence of enzyme protects its host from fluoroacetate toxicity in vivo 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 0.01
-
 acetyl-CoA wild-type, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.011
-
 acetyl-CoA mutant V23A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.012
-
 acetyl-CoA mutant T42C, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.029
-
 acetyl-CoA mutant F36A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.15
-
 fluoroacetyl-CoA mutant H76A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 0.16
-
 acetyl-CoA mutant T42S, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 4
-
 fluoroacetyl-CoA mutant T42A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 9
-
 fluoroacetyl-CoA mutant E50Q, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 18
-
 fluoroacetyl-CoA mutant F33A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 30
-
 fluoroacetyl-CoA mutant T42C, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 57
-
 fluoroacetyl-CoA mutant L26A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 300
-
 fluoroacetyl-CoA mutant T42S, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 370
-
 fluoroacetyl-CoA mutant F36A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 500
-
 fluoroacetyl-CoA mutant V23A, 25°C, pH 7.6 Streptomyces cattleya
3.1.2.29 50000
-
 fluoroacetyl-CoA wild-type, 25°C, pH 7.6 Streptomyces cattleya