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Literature summary for 4.4.1.1 extracted from

  • Hopwood, E.M.; Ahmed, D.; Aitken, S.M.
    A role for glutamate-333 of Saccharomyces cerevisiae cystathionine gamma-lyase as a determinant of specificity (2014), Biochim. Biophys. Acta, 1844, 465-472.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
E333A site-directed mutagenesis of the active-site residue, pH optimum 7.8-8.6, the mutant shows increased KM for L-cystathionine up to 17fold, and 2.5fold for O-acetyl-L-serine Saccharomyces cerevisiae
E333D site-directed mutagenesis of the active-site residue, pH optimum 7.2-8.0, the mutant shows increased KM for L-cystathionine up to 17fold, and 2.5fold for O-acetyl-L-serine Saccharomyces cerevisiae
E333Q site-directed mutagenesis of the active-site residue, pH optimum 7.6-8.4, the mutant shows increased KM for L-cystathionine up to 17fold, and 2.5fold for O-acetyl-L-serine Saccharomyces cerevisiae
E48A site-directed mutagenesis of the active-site residue, pH optimum 7.6-8.4 Saccharomyces cerevisiae
E48A/E333A site-directed mutagenesis of the active-site residues, pH optimum 8.4-9.2 Saccharomyces cerevisiae
E48D site-directed mutagenesis of the active-site residue, pH optimum 7.07.8 Saccharomyces cerevisiae
E48D/E333D site-directed mutagenesis of the active-site residues, pH optimum 7.6-8.4 Saccharomyces cerevisiae
E48Q site-directed mutagenesis of the active-site residue, pH optimum 7.4-8.2 Saccharomyces cerevisiae
E48Q/E333Q site-directed mutagenesis of the active-site residues, pH optimum 7.9-8.7 Saccharomyces cerevisiae
additional information pH optima of the mutant enzymes vary from wild-type enzyme, overview Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Saccharomyces cerevisiae
0.09
-
L-cysteine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae
0.11
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.18
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
0.4
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
0.4
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
0.5
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
0.5
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.7
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.7
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.71
-
L-cystathionine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae
0.76
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.87
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
0.9
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
0.9
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
1.2
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
1.4
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
1.5
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
1.5
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
1.8
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
2.2
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
2.5
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae
2.9
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
4.4
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
5.3
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
12
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
12
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
13
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-cystathionine + H2O Saccharomyces cerevisiae
-
L-cysteine + 2-oxobutanoate + NH3
-
r
L-cysteine + H2O Saccharomyces cerevisiae
-
H2S + pyruvate + NH3
-
?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-cystathionine + H2O
-
Saccharomyces cerevisiae L-cysteine + 2-oxobutanoate + NH3
-
r
L-cysteine + H2O
-
Saccharomyces cerevisiae H2S + pyruvate + NH3
-
?
additional information Glu333 of cystathionine gamma-lyase acts as a determinant of specificity, it interacts with the distal amine moiety of L-cystathionine, which is not present in the alternative substrate O-acetyl-L-serine. Catalytic efficiency of the yeast enzyme for alpha,gamma-elimination of O-succinyl-L-homoserine, which possesses a distal carboxylate, but lacks an amino group, is 300fold lower than that of the physiological L-cystathionine substrate and 260fold higher than that of L-Hcys, which lacks both distal polar moieties. Proposed polar contacts of the yCGL active site that interact with or are influenced by E48 and E333 overview Saccharomyces cerevisiae ?
-
?
O-acetyl-L-serine + H2O
-
Saccharomyces cerevisiae acetate + pyruvate + NH3
-
?

Synonyms

Synonyms Comment Organism
cgl
-
Saccharomyces cerevisiae
yCGL
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0035
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.0045
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
0.0047
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
0.0053
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
0.0058
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
0.0092
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
0.0097
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae
0.0102
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.0124
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
0.0142
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.029
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.09
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.11
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
0.15
-
L-cysteine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae
0.16
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
0.18
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
0.24
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
0.32
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
0.48
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.5
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.54
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.78
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
0.98
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
1.29
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
1.3
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
1.4
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
1.51
-
L-cystathionine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2 7.8 wild-type enzyme Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Saccharomyces cerevisiae

General Information

General Information Comment Organism
metabolism cystathionine gamma-lyase catalyzes the hydrolysis of L-cystathionine in the second step of the reverse transsulfuration pathway, which converts L-homocysteine to L-Cys Saccharomyces cerevisiae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0016
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
0.0019
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.0026
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
0.0028
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
0.0037
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
0.0038
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant wild-type enzyme and mutant E48D Saccharomyces cerevisiae
0.005
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
0.005
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.006
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
0.0088
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.01
-
O-acetyl-L-serine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.0169
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
0.036
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.06
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48D/E333D Saccharomyces cerevisiae
0.065
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
0.082
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
0.13
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333D Saccharomyces cerevisiae
0.21
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48A/E333A Saccharomyces cerevisiae
0.31
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48Q/E333Q Saccharomyces cerevisiae
0.49
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333Q Saccharomyces cerevisiae
0.71
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.72
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
0.8
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48A Saccharomyces cerevisiae
0.81
-
L-cysteine pH 7.2, 25°C, recombinant mutant E333A Saccharomyces cerevisiae
1.02
-
L-cysteine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
1.4
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48Q Saccharomyces cerevisiae
1.59
-
L-cystathionine pH 7.2, 25°C, recombinant mutant E48D Saccharomyces cerevisiae
1.7
-
L-cysteine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae
2.1
-
L-cystathionine pH 7.2, 25°C, recombinant wild-type enzyme Saccharomyces cerevisiae