Literature summary for 2.7.1.40 extracted from

Morgan, H.P.; McNae, I.W.; Nowicki, M.W.; Hannaert, V.; Michels, P.A.; Fothergill-Gilmore, L.A.; Walkinshaw, M.D.
Allosteric mechanism of pyruvate kinase from Leishmania mexicana uses a rock and lock model (2010), J. Biol. Chem., 285, 12892-12898.

Cloned(Commentary)

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 (Commentary)

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

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

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

Metals/Ions	Comment	Organism	Structure
K+	required	Leishmania mexicana
Mg2+	required	Leishmania mexicana

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + pyruvate	Leishmania mexicana	-	ADP + phosphoenolpyruvate	-	r

Organism

Organism	UniProt	Comment	Textmining
Leishmania mexicana	Q27686	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)	Leishmania mexicana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + pyruvate	-	Leishmania mexicana	ADP + phosphoenolpyruvate	-	r

Subunits

Subunits	Comment	Organism
homotetramer	enzyme structure in complex with ATP, oxalate, and fructose-2,6-bishosphate, overview	Leishmania mexicana

Synonyms

Synonyms	Comment	Organism
PYK	-	Leishmania mexicana

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Leishmania mexicana

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Leishmania mexicana

Cofactor

Cofactor	Comment	Organism	Structure
ADP	-	Leishmania mexicana
ATP	-	Leishmania mexicana

General Information

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