Purification and molecular properties of the AMP-activated pyruvate kinase from Escherichia coli.
The AMP-activated pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40) from Escherichia coli has been purified 200 times through a three-step procedure which gives a homogeneous preparation with a specific activity of 110. The enzyme appears to be a tetramer of molecular weight 190 000. Subunits (molecular weight 51 000) show a single amino-terminal amino acid (serine) and appear as a single band in polyacrylamide gel electrophoresis in sodium dodecyl sulphate. The enzyme crystallizes in conditions of reduced dielectric constant of the solvent in the pH range 6.5-7.5. Kinetic and regulatory properties of the purified enzyme are similar to those described for crude preparations of the enzyme.
D000596:Amino Acids / Q000032:analysis; D003460:Crystallization; D000242:Cyclic AMP / Q000494:pharmacology*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D005636:Fructosephosphates / Q000494:pharmacology; D006598:Hexosediphosphates / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011770:Pyruvate Kinase / Q000302:isolation & purification / Q000378:metabolism
D000596:Amino Acids; D005636:Fructosephosphates; D006598:Hexosediphosphates; D046911:Macromolecular Substances; D000242:Cyclic AMP; D011770:Pyruvate Kinase