Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), cloning in Escherichia coli strain XL-1 Blue | Leishmania mexicana |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type and mutant enzymes free and in complex with ligands ATP, oxalate, and fructose 2,6-bisphosphate, hanging drop vapour diffusion method, 0.0015 ml of 15 mg/ml protein in 20 mM TEA, pH 7.2, are mixed with 0.0015 ml of well solution composed of 10-16% PEG 8000, 20 mM TEA, pH 7.2, 50 mM MgCl2, 100 mM KCl, and 10-15% glycerol, 4°C or 17°C, 1week, X-ray diffraction structure determination and analysis | Leishmania mexicana |
General Stability | Organism |
---|---|
binding of frucctose 2,6-bisphophate plays the most important role in stabilizing the LmPYK tetramer in solution | Leishmania mexicana |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | required | Leishmania mexicana | |
Mg2+ | required | Leishmania mexicana |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + pyruvate | Leishmania mexicana | - |
ADP + phosphoenolpyruvate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leishmania mexicana | Q27686 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Leishmania mexicana |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + pyruvate | - |
Leishmania mexicana | ADP + phosphoenolpyruvate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | enzyme structure in complex with ATP, oxalate, and fructose-2,6-bishosphate, overview | Leishmania mexicana |
Synonyms | Comment | Organism |
---|---|---|
PYK | - |
Leishmania mexicana |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Leishmania mexicana |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Leishmania mexicana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ADP | - |
Leishmania mexicana | |
ATP | - |
Leishmania mexicana |
General Information | Comment | Organism |
---|---|---|
additional information | the transition between inactive T-state and active R-state is accompanied by a simple symmetrical 6o rigid body rocking motion of the A- and C-domain cores in each of the four subunits. Eight essential salt bridge locks form across the C-C interface providing tetramer rigidity with a coupled 7fold increase in reaction rate | Leishmania mexicana |
physiological function | the enzyme uses a rock and lock model allosteric mechanism, intersubunit interactions on the A-A and C-C interfaces strongly influence the allosteric effect whereas mutations affecting the intrasubunit A-C interface are less sensitive, overview. Conformational changes coupled with effector binding correlate with loss of flexibility and increase in thermal stability providing a general mechanism for allosteric control | Leishmania mexicana |