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Literature summary for 4.4.1.13 extracted from

  • Jaworski, A.F.; Aitken, S.M.
    Exploration of the six tryptophan residues of Escherichia coli cystathionine beta-lyase as probes of enzyme conformational change (2013), Arch. Biochem. Biophys., 538, 138-144.
    View publication on PubMed
Search for more literature for 4.4.1.13

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain ER1821 metC::cat Escherichia coli

Protein Variants

Protein Variants Comment Organism
W131F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W188F/W230F/W276F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W188F/W230F/W276F/W300F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W188F/W230F/W276F/W340F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W188F/W230F/W300F/W340F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W188F/W276F/W300F/W340F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W230F/W276F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W230F/W276F/W300F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W230F/W276F/W300F/W340F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W131F/W230F/W276F/W340F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W188F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by 4.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W188F/W230F/W276F/W300F/W340F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W230F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W276F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W300F site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7°C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme Escherichia coli
W340F site-directed mutagenesis, 8fold ncrease in KM for L-cystathionine compared to the wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Escherichia coli
0.101
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F Escherichia coli
0.116
-
 L-cystathionine pH 8.5, 25°C, recombinant wild-type enzyme Escherichia coli
0.121
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W188F Escherichia coli
0.131
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W276F Escherichia coli
0.148
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W300F Escherichia coli
0.161
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W300F Escherichia coli
0.171
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131FF/W230FF/W276F Escherichia coli
0.19
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W230F Escherichia coli
0.27
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F Escherichia coli
0.31
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W300F Escherichia coli
0.91
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W340F Escherichia coli
1
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W340F Escherichia coli
1.05
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W276F/W300F/W340F Escherichia coli
1.26
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W300F/W340F Escherichia coli
1.8
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W300F/W340F Escherichia coli
1.8
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W340F Escherichia coli
2
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W188F/W230F/W276F/W300F/W340F Escherichia coli
2.1
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W300F/W340F Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-cystathionine + H2O Escherichia coli
-
 L-homocysteine + pyruvate + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P06721
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain ER1821 metC::cat by nickel affinity chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-cystathionine + H2O
-
 Escherichia coli L-homocysteine + pyruvate + NH3
-
 ?

Synonyms

Synonyms Comment Organism
CBL
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 Tm values of wild-type and mutant enzymes, overview Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
25.4
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F Escherichia coli
31.7
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W300F Escherichia coli
35.1
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W276F/W300F/W340F Escherichia coli
38
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W300F/W340F Escherichia coli
38.1
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W188F Escherichia coli
40
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W340F Escherichia coli
42
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W188F/W230F/W276F/W300F/W340F Escherichia coli
42.5
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W300F Escherichia coli
45.3
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F Escherichia coli
46
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W300F/W340F Escherichia coli
54
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W300F/W340F Escherichia coli
54.5
-
 L-cystathionine pH 8.5, 25°C, recombinant wild-type enzyme Escherichia coli
54.5
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131FF/W230FF/W276F Escherichia coli
55.9
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W230F Escherichia coli
60.4
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W276F Escherichia coli
60.7
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W340F Escherichia coli
65
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W340F Escherichia coli
74
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W300F Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
 Escherichia coli

General Information

General Information Comment Organism
additional information conserved residues W188 and W340 are situated at the core of the domain interface that forms the active-site cleft, W188 is a useful probe of subtle conformational changes at the domain interface and active site. The active site, containing the pyridoxal 5'-phosphate cofactor is situated at the interface between the catalytic (residues 61-256) and C-terminal (residues 257-395) domains. The N-terminal domain (residues 1-60) contributes to the active site of the neighboring subunit Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
18
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W300F/W340F Escherichia coli
21
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W188F/W230F/W276F/W300F/W340F Escherichia coli
23
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W340F Escherichia coli
25
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W300F/W340F Escherichia coli
33
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W276F/W300F/W340F Escherichia coli
43
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W300F/W340F Escherichia coli
66
-
 L-cystathionine pH 8.5, 25°C, recombinant mutants W340F and W131F/W230F/W276F/W340F Escherichia coli
94
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F Escherichia coli
213
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W188F/W230F/W276F/W300F Escherichia coli
240
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F/W230F/W276F/W300F Escherichia coli
260
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W300F Escherichia coli
300
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W230F Escherichia coli
315
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W188F Escherichia coli
320
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131FF/W230FF/W276F Escherichia coli
450
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W131F Escherichia coli
460
-
 L-cystathionine pH 8.5, 25°C, recombinant mutant W276F Escherichia coli
470
-
 L-cystathionine pH 8.5, 25°C, recombinant wild-type enzyme Escherichia coli