Improved procedures for the synthesis of phosphomevalonate and for the assay and purification of pig liver phosphomevalonate kinase

• Published in: Biochimica et biophysica acta Vol. 839; no. 1; pp. 83 - 89
• Main Authors: Lee, Choy Soong, O'Sullivan, William J.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 29.03.1985; Elsevier; North-Holland
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cations, Divalent; Electrophoresis; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Liver - enzymology; Methods; Mevalonic Acid - analogs & derivatives; Mevalonic Acid - chemical synthesis; Molecular Weight; Phosphomevalonate kinase; Phosphomevalonate synthesis; Phosphotransferases (Phosphate Group Acceptor); Phosphotransferases - analysis; Phosphotransferases - metabolism; Pig liver; Substrate Specificity; Swine; Transferases; Pig liver; Mes; Phosphomevalonate synthesis; Phosphomevalonate kinase; Hepes; Assay; Vertebrata; Mammalia; Purification; Enzyme; Liver; Artiodactyla; Catalyst activity; Ungulata; Pig
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(85)90184-9

An improved procedure for the synthesis of phosphomevalonate using excess free ATP4−, and phenyl agarose to remove contaminating nucleotides, is described. A high-voltage electrophoresis assay, which separates phosphomevalonate from mevalonate 5-diphosphate at pH 3.5, was developed for the assay of phosphomevalonate kinase (ATP:5-phosphomevalonate phosphotransferase, EC 2.7.4.2). High-voltage electrophoresis, at pH 5, could also be used for the separation of mevalonate 5-diphosphate from isopentenyl diphosphate. An alternative method for the purification of phosphomevalonate kinase from pig liver was also developed. The high-voltage electrophoresis assay was used to reasses the metal ion and nucleotide specificity of the pig liver phosphomevalonate kinase. ATP could be partially replaced by ITP and GTP and poorly by CTP and UTP. Apparent activation of the enzyme by free ATP4− was observed as found for mevalonate kinase (C.S. Lee and W.J. O'Sullivan (1983) Biochim. Biophys. Acta 747, 215–224).