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Analysis Note

Protein determined by biuret.

Application

Pyruvate kinase is an enzyme involved in glycolysis and is involved in 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.

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

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

Molecular Weight: 237 kDa and exists as a tetramer of four equal subunits of molecular weight 57 kDa.Isoelectric Point: 7.6Optimal 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.

Physical form

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

Unit Definition

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