Subunit structure and some properties of pyruvate kinase of Neurospora.
Pyruvate kinase isolated from Neurospora and purified to homogeneity has been shown to be a tetramer of molecular weight around 242 000 by gel filtration studies and 239 000 daltons by sedimentation equilibrium measurements. The monomer produced by treatment with guanidine hydrochloride is found to be 51 000-52 000 daltons by sedimentation equilibrium studies; a molecular weight of 62 000 was determined for the monomer generated by SDS treatment by electrophoresis in SDS-polyacrylamide gels. The enzyme has an isoelectric point of 6.35-6.41; Substrate saturation kinetics of PEP show a variable extent of cooperativity depending upon the buffer ions employed in the assay. ADP is the most effective phosphoryl group acceptor, GDP and IDP being poor substitutes. A divalent cation, Mg-2+, is required for activity. At low concentrations, Ca-2+ acts as an activator of pyruvate kinase but it is inhibitory  at high concentrations. Fructose 1,6-diphosphate is the most potent allosteric activator, fructose 6-phosphate being next in order of effectiveness. Valine is a powerful inhibitor. Phenylalanine, tyrosine, and tryptophan are without any effect individually, but their simultaneous presence results in a considerable activation. Alanine does not affect this enzyme appreciably.
D000494:Allosteric Regulation; D002118:Calcium / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005636:Fructosephosphates / Q000494:pharmacology; D005976:Glutaral; D006598:Hexosediphosphates / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D011770:Pyruvate Kinase / Q000302:isolation & purification / Q000378:metabolism; D014461:Ultracentrifugation; D014633:Valine / Q000494:pharmacology
D005636:Fructosephosphates; D006598:Hexosediphosphates; D046911:Macromolecular Substances; D011770:Pyruvate Kinase; D014633:Valine; D008274:Magnesium; D002118:Calcium; D005976:Glutaral