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Key Documents

P7768

Sigma-Aldrich

Pyruvate Kinase from rabbit muscle

Type VII, buffered aqueous glycerol solution, 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:
MDL number:
UNSPSC Code:
12352204
eCl@ss:
32160410
NACRES:
NA.54

type

Type VII

Quality Level

form

buffered aqueous glycerol solution

specific activity

350-600 units/mg protein

mol wt

237 kDa

concentration

2.0-20.0 mg/mL

foreign activity

lactic dehydrogenase and creatine phosphokinase ≤0.01%
phosphoglucomutase and myokinase ≤0.05%

storage temp.

2-8°C

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General description

Research Area: Cell Signaling
Pyruvate kinase plays a role in regulating cell metabolism. There are four pyruvate kinase isoforms in mammals (PKM1, PKM2, PKR, PKL). Mammalian pyruvate kinase is a tetrameric protein composed of identical subunits, arranged in a dimer-of-dimers configuration. Each monomer contains one active site and consists of three main domains- designated A, B, and C-along with a small N-terminal domain. The M2 isoform of pyruvate kinase (PKM2) supports anabolic metabolism and is expressed in cancer and normal tissue.

Application

Pyruvate Kinase from rabbit muscle has been used as a component in refolding buffer for malate dehydrogenase (MDH) assay. It has also been used as a supplement in assay buffer for phosphoglycerate mutases (PGM) enzyme assays.
Pyruvate kinase from Sigma has been used in the preparation of cell free extract from BL21 CP strain of E. coli culture for highly productive cell-free protein expression. The enzyme has been used to assay the pyruvate kinase activity by incubating with eluted DAPk (Death-associated protein kinase), a serine/threonine kinase that binds and activates pyruvate kinase.

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 catalyzes the transfer of a phosphate group from phosphoenolpyruvate (PEP) to ADP. This transfer yields one molecule of pyruvate and one molecule of ATP. 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
Pyruvate kinase is an enzyme involved in glycolysis and gluconeogenesis. Product P7768 is from rabbit muscle and is provided in a buffered aqueous glycerol solution and has been used in PGM enzyme assays to determine phosphoglycerate mutase activity. Pyruvate kinase is also used to study pyruvate kinase (PK) deficiency

Unit Definition

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

Physical form

Solution in 50% glycerol containing 0.01 M phosphate, pH 7.0

Analysis Note

Protein determined by biuret.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


Certificates of Analysis (COA)

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James F Zawada
Methods in molecular biology (Clifton, N.J.), 805, 31-41 (2011-11-19)
Crude cell-free extracts are useful tools for investigating biochemical phenomena and exploiting complex enzymatic processes such as protein synthesis. Extracts derived from E. coli have been used for over 50 years to study the mechanism of protein synthesis. In addition
I Mor et al.
Oncogene, 31(6), 683-693 (2011-07-05)
Death-associated protein kinase (DAPk), a multi-domain serine/threonine kinase, regulates numerous cell death mechanisms and harbors tumor suppressor functions. In this study, we report that DAPk directly binds and functionally activates pyruvate kinase M2 (PKM2), a key glycolytic enzyme, which contributes
Structural-functional diversity of malaria parasite's PfHSP70-1 and PfHSP40 chaperone pair gives an edge over human orthologs in chaperone-assisted protein folding
Anas M, et al.
The Biochemical Journal, 477(18), 3625-3643 (2020)
Hager Souabni et al.
Communications biology, 4(1), 493-493 (2021-04-24)
Tripartite efflux pumps built around ATP-binding cassette (ABC) transporters are membrane protein machineries that perform vectorial export of a large variety of drugs and virulence factors from Gram negative bacteria, using ATP-hydrolysis as energy source. Determining the number of ATP
Studies on pyruvate kinase (PK) deficiency. II. Electrophoretic, kinetic and immunological studies on pyruvate kinase of erythrocytes and other tissues.
K Imamura et al.
Journal of biochemistry, 74(6), 1165-1175 (1973-12-01)

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