Application | Comment | Organism |
---|---|---|
pharmacology | the enzyme is a candidate chemotherapeutic target | Clonorchis sinensis |
Cloned (Comment) | Organism |
---|---|
gene CLF_107872, DNA and amino acid sequence determination and analysis, sequence comarisons and phylogenetic analysis, the coding region of the cDNA of D1D2 is cloned into the pMAL-c2 vector and expressed as maltose binding protein fusion protein in Escherichia coli strain TB1, quantitative real-time PCR enzyme expression analysis. Recombinant expression of N-terminally GST-tagged enzyme D1 and D2 mutants in Escherichia coli strain BL21(DE3) pLysS | Clonorchis sinensis |
Protein Variants | Comment | Organism |
---|---|---|
H61A | site-directed mutagenesis of a domain 2 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
H61A | site-directed mutagenesis of TKD1D2 in D2 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
I60A | site-directed mutagenesis of a domain 1 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
I60A | site-directed mutagenesis of TKD1D2 in D1 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
I63A | site-directed mutagenesis of a domain 2 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
I63A | site-directed mutagenesis of TKD1D2 in D2 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
R58A | site-directed mutagenesis of a domain 1 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
R58A | site-directed mutagenesis of TKD1D2 in D1 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y84A | site-directed mutagenesis of a domain 1 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y84A | site-directed mutagenesis of TKD1D2 in D1 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y84R | site-directed mutagenesis of a domain 1 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y84R | site-directed mutagenesis of TKD1D2 in D1 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y87A | site-directed mutagenesis of a domain 2 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y87A | site-directed mutagenesis of TKD1D2 in D2 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y87R | site-directed mutagenesis of a domain 2 residue, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
Y87R | site-directed mutagenesis of TKD1D2 in D2 region, the enzyme mutant shows altered kinetics and reduced activity compared to the wild-type enzyme | Clonorchis sinensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | comparisons of wild-type and mutant enzyme kinetics, overview | Clonorchis sinensis | |
0.35 | - |
Taurocyamine | recombinant domain D1, pH 8.0, 25°C | Clonorchis sinensis | |
0.46 | - |
ATP | recombinant domain D1, pH 8.0, 25°C | Clonorchis sinensis | |
0.48 | - |
Taurocyamine | recombinant domain D2, pH 8.0, 25°C | Clonorchis sinensis | |
0.49 | - |
Taurocyamine | recombinant full-length enzyme, pH 8.0, 25°C | Clonorchis sinensis | |
0.75 | - |
ATP | recombinant domain D2, pH 8.0, 25°C | Clonorchis sinensis | |
0.78 | - |
ATP | recombinant full-length enzyme, pH 8.0, 25°C | Clonorchis sinensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Clonorchis sinensis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
39720 | - |
sequence calculation, D2 domain | Clonorchis sinensis |
40570 | - |
sequence calculation, D1 domain | Clonorchis sinensis |
42000 | - |
- |
Clonorchis sinensis |
80000 | - |
- |
Clonorchis sinensis |
80270 | - |
sequence calculation, full-length enzyme | Clonorchis sinensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + taurocyamine | Clonorchis sinensis | - |
ADP + N-phosphotaurocyamine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clonorchis sinensis | G7YHA8 | gene CLF_107872 | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme coding region as maltose binding protein fusion protein from Escherichia coli strain TB1 by amylose affinity chromatography too homogeneity. Recombinant N-terminally GST-tagged enzyme D1 and D2 mutants from Escherichia coli strain BL21(DE3) pLysSby glutathione affinity chromatography, the GST-tag is cleaved off by thrombin treatment | Clonorchis sinensis |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
egg | intra-uterine | Clonorchis sinensis | - |
integument | - |
Clonorchis sinensis | - |
intestine | - |
Clonorchis sinensis | - |
additional information | enzyme expression is distributed in locomotive and reproductive organs of adult worms. The enzyme is stably expressed in both the adult and metacercariae stages, quantitative real time PCR expression analysis, overview. Immunohistochemic analysis | Clonorchis sinensis | - |
seminal vesicle | - |
Clonorchis sinensis | - |
sperm | - |
Clonorchis sinensis | - |
sucker | oral and ventral suckers | Clonorchis sinensis | - |
testis | - |
Clonorchis sinensis | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + taurocyamine | - |
Clonorchis sinensis | ADP + N-phosphotaurocyamine | - |
? | |
ATP + taurocyamine | high substrate specificity for taurocyamine | Clonorchis sinensis | ADP + N-phosphotaurocyamine | - |
? | |
additional information | residues R58, I60 and Y84 of domain 1, and H60, I63 and Y87 of domain 2 participate in binding taurocyamine. Recombinant CsTK enzyme protein has low sensitivity and specificity toward Clonorchis sinensis and platyhelminth-infected human sera on ELISA | Clonorchis sinensis | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 42000, SDS-PAGE, recombinant detagged D1 domain, SDS-PAGE, 1 * 80000, recombinant full-length enzyme, SDS-PAGE | Clonorchis sinensis |
Synonyms | Comment | Organism |
---|---|---|
CsTK | - |
Clonorchis sinensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Clonorchis sinensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.5 | - |
ATP | recombinant domain D2, pH 8.0, 25°C | Clonorchis sinensis | |
4.5 | - |
Taurocyamine | recombinant domain D2, pH 8.0, 25°C | Clonorchis sinensis | |
17.78 | - |
ATP | recombinant full-length enzyme, pH 8.0, 25°C | Clonorchis sinensis | |
17.78 | - |
Taurocyamine | recombinant full-length enzyme, pH 8.0, 25°C | Clonorchis sinensis | |
22.59 | - |
ATP | recombinant domain D1, pH 8.0, 25°C | Clonorchis sinensis | |
22.59 | - |
Taurocyamine | recombinant domain D1, pH 8.0, 25°C | Clonorchis sinensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Clonorchis sinensis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Clonorchis sinensis |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Clonorchis sinensis | sequence calculation, D2 domain | - |
7.39 |
Clonorchis sinensis | sequence calculation, full-length enzyme | - |
7.88 |
Clonorchis sinensis | sequence calculation, D1 domain | - |
8.09 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the highly conserved phosphagen (PK) kinase family of enzymes. The polypeptide is homologous to taurocyamine kinase (TK) of the invertebrate animals and consists of two contiguous domains. Clonorchis sinensis CsTK gene consists of 13 exons intercalated with 12 introns. This suggests an evolutionary pathway originating from an arginine kinase gene group, and distinguishes annelid taurocyamine kinase from the general creatine kinase phylogenetic group | Clonorchis sinensis |