Literature summary for 2.7.1.40 extracted from

Zhong, W.; Li, K.; Cai, Q.; Guo, J.; Yuan, M.; Wong, Y.H.; Walkinshaw, M.D.; Fothergill-Gilmore, L.A.; Michels, P.A.M.; Dedon, P.C.; Lescar, J.
Pyruvate kinase from Plasmodium falciparum structural and kinetic insights into the allosteric mechanism (2020), Biochem. Biophys. Res. Commun., 532, 370-376 .

Activating Compound

Activating Compound	Comment	Organism	Structure
glucose 6-phosphate	G6P, an activator increasing the apparent maximal velocity of isozyme PYK-I, 1.5fold activation at 5 mM without affecting the affinity and cooperativity towards the PEP substrate. The binding of glucose 6-phosphate and oxalate, which potentially lock the enzyme in its active state, increase the thermal stability of the enzyme. PfPYK might be a V-type allosteric enzyme with respect to G6P. In silico docking of the activator G6P to the canonical effector site, the phosphate group of G6P forms a number of favorable interactions with the PO4-2 motif	Plasmodium falciparum
additional information	no effects by fructose 1,6-bisphosphate or fructose 2,6-bisphosphate on the enzyme activity	Plasmodium falciparum

Cloned(Commentary)

Cloned (Comment)	Organism
gene PF3D7_1037100, sequence comparisons, recombinant expression of GST-tagged enzyme in Escherichia coli. It is likely that the GST-tag partially hinders or affects conformational changes occurring in the PfPYK-I tetramer, which are crucial for allosteric regulation	Plasmodium falciparum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified isozyme PYK-I in complex with substrate analogue oxalate, activator glucose 6-phosphate, and product ATP, hanging drop vapour diffusion method, mixing of 0.001 ml of protein solution and 500 nl of 20 mM ligand solution, with 0.0015 ml of reservoir solution containing 12% PEG 8000, 10-20% glycerol, 50 mM TEA, pH 7.2, 100 mM KCl, and 50 mM MgCl2, and equilibration against 1 ml of reservoir solution, X-ray diffraction structure determination and analysis, molecular replacement using the structure obtained from the deposited T-state PfPYK (PDB ID 3KHD) as template, molecular modelling	Plasmodium falciparum

Crystallization (Comment)

Organism

purified isozyme PYK-I in complex with substrate analogue oxalate, activator glucose 6-phosphate, and product ATP, hanging drop vapour diffusion method, mixing of 0.001 ml of protein solution and 500 nl of 20 mM ligand solution, with 0.0015 ml of reservoir solution containing 12% PEG 8000, 10-20% glycerol, 50 mM TEA, pH 7.2, 100 mM KCl, and 50 mM MgCl2, and equilibration against 1 ml of reservoir solution, X-ray diffraction structure determination and analysis, molecular replacement using the structure obtained from the deposited T-state PfPYK (PDB ID 3KHD) as template, molecular modelling

Plasmodium falciparum

Inhibitors

Inhibitors	Comment	Organism
citrate	citrate at 2 mM inhibits GST-tagged PfPYK activity by over 90%, citrate slightly decreases the affinity for the PEP substrate, with no obvious change in the apparent kcat	Plasmodium falciparum
additional information	no effects by fructose 1,6-bisphosphate or fructose 2,6-bisphosphate on the enzyme activity	Plasmodium falciparum
oxalate	the binding of glucose 6-phosphate and oxalate, which potentially lock the enzyme in its active state, increase the thermal stability of the enzyme	Plasmodium falciparum
suramin	-	Plasmodium falciparum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	allosteric mechanism and structural changes, overview	Plasmodium falciparum
0.00017	-	ADP	pH and temperature not specified in the publication	Plasmodium falciparum
0.00039	-	ADP	in presence of inhibitor suramin, pH and temperature not specified in the publication	Plasmodium falciparum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ADP + phosphoenolpyruvate	Plasmodium falciparum	-	ATP + pyruvate	-	?

Organism

Organism	UniProt	Comment	Textmining
Plasmodium falciparum	C6KTA4	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ADP + phosphoenolpyruvate	-	Plasmodium falciparum	ATP + pyruvate	-	?

Subunits

Subunits	Comment	Organism
homotetramer	4 * 55000, PYK-I, SDS-PAGE	Plasmodium falciparum
More	structure-function analysis, overview	Plasmodium falciparum

Synonyms

Synonyms	Comment	Organism
PfPYK	-	Plasmodium falciparum
PYK	-	Plasmodium falciparum
PYK-I	-	Plasmodium falciparum

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	the binding of glucose 6-phosphate and oxalate, which potentially lock the enzyme in its active state, increase the thermal stability of the enzyme	Plasmodium falciparum

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.149	-	pH and temperature not specified in the publication	Plasmodium falciparum	oxalate

General Information

General Information	Comment	Organism
additional information	comparisons of isozyme PYK-I structures in the active R-state and inactive T-state reveal a rock-and-lock allosteric mechanism regulated by rigid-body rotations of each subunit in the tetramer. It is likely that the GST-tag on the recombinant enzyme partially hinders or affects conformational changes occurring in the PfPYK-I tetramer, which are crucial for allosteric regulation. Structure-function analysis, overview	Plasmodium falciparum