P7768

Pyruvate Kinase from rabbit muscle

Type VII, buffered aqueous glycerol solution, 350-600 units/mg protein

• Synonym(s): ATP:pyruvate 2-O-phosphotransferase, PK
• CAS Number: 9001-59-6
• Enzyme Commission number: 2.7.1.40 (BRENDA, IUBMB)
• MDL number: MFCD00081946
• UNSPSC Code: 12352204
• eCl@ss: 32160410
• NACRES: NA.54

Properties

• type: Type VII
• Quality Level: 300
• 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

Description

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 K_M 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.

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves