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P9136

Sigma-Aldrich

Pyruvate Kinase from rabbit muscle

Type III, lyophilized powder, 350-600 units/mg protein

Synonym(s):

ATP:pyruvate 2-O-phosphotransferase, PK

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About This Item

CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
eCl@ss:
32160410
NACRES:
NA.54

biological source

rabbit muscle

Quality Level

type

Type III

form

lyophilized powder

specific activity

350-600 units/mg protein

mol wt

237 kDa

foreign activity

lactic dehydrogenase, creatine phosphokinase, phosphoglucomutase, and myokinase ≤0.01%

storage temp.

−20°C

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General description

Pyruvate Kinase (PK) is a glycolysis enzyme and has four isozymes L, R, M1, and M2. The L isozyme is localized in gluconeogenic tissues, particularly in the liver. Whereas the R and M1 are localized in the adult skeletal muscles, heart, brain and erythrocytes, respectively. M2 is localized in the nucleus of the cells.

Application

Pyruvate Kinase from rabbit muscle has been used to convert adenosine diphosphate (ADP) to adenosine triphosphate (ATP) in Pseudomonas putida cells.
Pyruvate kinase has been used in plant spectrophotometric assays to measure ATP hydrolysis . Pyruvate kinase is also used to study pyruvate kinase (PK) deficiency .

Biochem/physiol Actions

Molecular Weight: 237 kDa and exists as a tetramer of four equal subunits of molecular weight 57 kDa.
Isoelectric Point: 7.6
Optimal pH: ∼7.5
Optimal Temperature: 25°C
ΕA280 = 0.54 for 1 mg(p)/ml, 1 cm path
Reported KM values are ATP (0.86 mM), pyruvate (10 mM), ADP (0.3 mM), and PEP (0.07 mM) in Tris buffer at pH 7.4 and 30 °C. Pyruvate kinase is highly specific for phosphoenolpyruvate, but can utilize other dinucleotide triphosphates as substrates in place of ATP including GTP, ITP, dATP, UTP, and CTP.
Pyruvate kinase (PK) catalyzes an important process of transferring phosphate group from phosphoenolpyruvate (PEP) to adenosine diphosphate (ADP). This reaction results in the conversion of PEP to pyruvate and adenosine triphosphate (ATP). Pyruvate kinase plays a major role in glycolysis and gluconeogenesis. High levels of pyruvate kinase M2 (PKM2) inhibits cell proliferation and tumor growth. Deficiency of PK in red blood cells (RBC) leads to non-spherocytic hemolytic anaemia. In human erythrocytes, deficiency of PK induces a protective effect against Plasmodium falciparum.

Unit Definition

One unit will convert 1.0 μmole of phospho(enol)pyruvate to pyruvate per min at pH 7.6 at 37 °C.

Analysis Note

Protein determined by biuret.

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


Certificates of Analysis (COA)

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Red cell pyruvate kinase deficiency: molecular and clinical aspects
Zanella A, et al.
British Journal of Haematology, 130(1), 11-25 (2005)
Pyruvate kinase deficiency and malaria
Ayi K, et al.
The New England Journal of Medicine, 358(17), 1805-1810 (2008)
Anoxic metabolism and biochemical production in Pseudomonas putida F1 driven by a bioelectrochemical system
Lai B, et al.
Biotechnology for Biofuels, 9(1), 39-39 (2016)
[25] Pyruvate kinase isozymes from rat
Methods in Enzymology, 90, 150-165 (1982)
S P Robinson et al.
Plant physiology, 88(4), 1008-1014 (1988-12-01)
Ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco) activase protein was purified from spinach leaves by ammonium sulfate precipitation and ion exchange fast protein liquid chromatography. This resulted in 48-fold purification with 70% recovery of activity and yielded up to 18 milligrams of rubisco

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