Cloned (Comment) | Organism |
---|---|
expression of soluble wild-type and mutant enzymes in HEK-293 cells | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
purified wild-type enzyme and mutant S203A in complex with substrates acetylcholine, acetylthiocholine, succinyldicholine, and butyrylthiocholine, and two nonhydrolyzable substrate analogues (m-N,N,N-trimethylammonio)-trifluoroacetophenone and 4-ketoamyltrimethylammonium, by hanging drop vapor diffusion at 4°C, using a protein-to-well solution ratio of 1:1, with well solution containing 2532% v/v P550MME or P600 in either 60-100 mM HEPES or sodium acetate, pH 6.5-pH 8.0, he TMTFA-mAChE complex is formed in solution using apo-mAChE at 5.0 mg/ml, and a 3-fold molar excess of the ligand over the enzyme, concentrated prior to crystallization, the other seven complexes are generated by crystal soaking, carried out at 4 °C in 0.02 ml sitting drops made of well solution supplemented with the 250 mM ligand and polyethylene glycol up to 35%, X-ray diffraction structure determination and anaylsis at 2.05-2.65 A resolution | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
S203A | site-directed mutagenesis of the catalytic site residue, inactive mutant | Mus musculus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
4-ketoamyltrimethylammonium | binding structure analysis | Mus musculus | |
choline | product inhibition, binding structure analysis | Mus musculus | |
m-(N,N,N-trimethylammonio)-trifluoroacetophenone | binding structure analysis | Mus musculus | |
succinyldicholine | substrate inhibition, binding structure analysis | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | ligand binding kinetics, overview | Mus musculus | |
0.046 | - |
acetylthiocholine | pH 7.0, 22°C, recombinant wild-type enzyme | Mus musculus | |
0.093 | - |
butyrylthiocholine | pH 7.0, 22°C, recombinant wild-type enzyme | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetylcholine + H2O | Mus musculus | - |
choline + acetate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | P21836 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant soluble wild-type and mutant enzymes from HEK-293 cells by by affinity chromatography with elution using either propidium diiodide or SCh dichloride, followed by dialysis | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetylcholine + H2O | - |
Mus musculus | choline + acetate | - |
? | |
acetylthiocholine + H2O | - |
Mus musculus | thiocholine + acetate | - |
? | |
butyrylthiocholine + H2O | - |
Mus musculus | thiocholine + butyrate | - |
? | |
additional information | substrate and product trafficking through the active center gorge of the enzyme analyzed by crystallography and equilibrium binding, binding structures, overview | Mus musculus | ? | - |
? | |
succinyldicholine + H2O | - |
Mus musculus | 2 choline + succinate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AChE | - |
Mus musculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Mus musculus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.021 | - |
succinyldicholine | pH 7.0, 22°C, recombinant wild-type enzyme | Mus musculus | |
0.023 | - |
4-ketoamyltrimethylammonium | pH 7.0, 22°C, recombinant wild-type enzyme | Mus musculus | |
1.4 | - |
choline | pH 7.0, 22°C, recombinant wild-type enzyme | Mus musculus |