Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain ER1821 metC::cat | Escherichia coli |
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 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-cystathionine + H2O | Escherichia coli | - |
L-homocysteine + pyruvate + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P06721 | - |
- |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-cystathionine + H2O | - |
Escherichia coli | L-homocysteine + pyruvate + NH3 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CBL | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Escherichia coli |
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 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 |