Literature summary for 2.7.1.40 extracted from

Cook, W.J.; Senkovich, O.; Aleem, K.; Chattopadhyay, D.
Crystal structure of Cryptosporidium parvum pyruvate kinase (2012), PLoS ONE, 7, e46875.

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli strain BL21(DE3)	Cryptosporidium parvum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant untagged enzyme in apoform or complexed with the non-hydrolyzable ATP analogue adenyl-5'-yl imidodiphosphate, hanging drop vapor diffusion technique, 4°C, mixing of 7 mg/ml protein in 0.65 M ammonium sulfate and 0.1 M sodium acetate, pH 4.0, and with 3 mM adenyl-5'-yl imidodiphosphate, 5 mM magnesium chloride, and 5 mM pyruvic acid for the enzyme complex, X-ray diffraction structure determination and analysis at 2.5 A resolution, molecular replacement	Cryptosporidium parvum

Crystallization (Comment)

Organism

purified recombinant untagged enzyme in apoform or complexed with the non-hydrolyzable ATP analogue adenyl-5'-yl imidodiphosphate, hanging drop vapor diffusion technique, 4°C, mixing of 7 mg/ml protein in 0.65 M ammonium sulfate and 0.1 M sodium acetate, pH 4.0, and with 3 mM adenyl-5'-yl imidodiphosphate, 5 mM magnesium chloride, and 5 mM pyruvic acid for the enzyme complex, X-ray diffraction structure determination and analysis at 2.5 A resolution, molecular replacement

Cryptosporidium parvum

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Cryptosporidium parvum
sulfate	each monomer in the asymmetric unit of CpPyK binds two sulfate ions at equivalent positions, one in the C-domain and the other at the interface of the A and C domains, binding structure, overview. The sulfate ion in the C-domain occupies a position corresponding to the 6-phosphate of the effector molecule in different PyKs	Cryptosporidium parvum

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Cryptosporidium parvum	Q5CSM7	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinnat enzyme from Eschericchia coli strain BL21(DE3) by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration	Cryptosporidium parvum

Substrates and Products (Substrate)

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

Subunits

Subunits	Comment	Organism
tetramer	three-dimensional structure determination and analysis, structure comparisons, overview. Each CpPyK monomer consists of four domains: N (residues 23-32), A (residues 42-112 and 212-389), B (residues 113-211) and C (residues 390-526). The A-domain constitutes the central part of the molecule and forms a parallel (alphaa/beta)8 barrel. The B-domain contains nine beta strands that form an antiparallel beta-barrel. The active site is located at the interface of the A and B domains, and residues from both domains participate in substrate binding. The C-domain is composed of five beta strands surrounded by five alpha-helices. The allosteric site for binding the effector molecule is located in the C-domain. The A domains of the two monomers A and B form the major interface	Cryptosporidium parvum

Synonyms

Synonyms	Comment	Organism
PYK	-	Cryptosporidium parvum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at	Cryptosporidium parvum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5.5	7.5	activity range	Cryptosporidium parvum

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Cryptosporidium parvum

General Information

General Information	Comment	Organism
additional information	the partially closed active site structure contains an alpha6' helix that unwinds and assumes an extended conformation, a glycerol molecule is located near the gamma-phosphate site of ATP. A sulfate ion is found at a site that is occupied by a phosphate of the effector molecule	Cryptosporidium parvum
physiological function	pyruvate kinase of Cryptosporidium parvum is exceptional among known enzymes of protozoan origin in that it exhibits no allosteric property in the presence of commonly known effector molecules, mainly phosphosugars, due to blockage of the effector binding site by a sulfate ion, overview	Cryptosporidium parvum