title: A fructose bisphosphate activated lactate dehydrogenase in the liver fluke Fasciola hepatica.
abstract: Lactate dehydrogenase of Fasciola hepatica showed typical Michaelis-Menten kinetics at pH 7.2, with respect to pyruvate. Addition of physiological levels of fructose bisphosphate activated the enzyme at all substrate concentrations tested; the response to this effector being hyperbolic in nature. As well as depending upon the fructose bisphosphate concentration, the Vmax and Km are modified by different buffers. The degree of activation is much greater using Tris-HCl than phosphate buffer. The pH optimum occurs at pH 6.5 whether using physiological levels of substrate in the presence or absence of fructose bisphosphate, or high levels of substrate. Of the potential effectors tested, significant inhibition was shown by the nucleoside triphosphates, especially ATP. The importance of this inhibition, coupled with the activation by fructose bisphosphate is discussed. Fasciola hepatica lactate dehydrogenase is unusual in that it does not catalyse the reverse reaction to any measurable extent. That is, lactate oxidation is negligible unless the effector fructose bisphosphate is present. Use was made of this fact to visualise the isoenzymes of lactate dehydrogenase separated by polyacrylamide disc gel electrophoresis. Five isoenzyme bands became apparent when stained in this manner.
mesh_terms: D000818:Animals; D004789:Enzyme Activation; D005210:Fasciola hepatica / Q000201:enzymology; D005211:Fascioliasis / Q000201:enzymology* / Q000469:parasitology; D005635:Fructosediphosphates / Q000502:physiology*; D006598:Hexosediphosphates / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D011773:Pyruvates / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D005635:Fructosediphosphates; D006598:Hexosediphosphates; D007527:Isoenzymes; D011773:Pyruvates; D007770:L-Lactate Dehydrogenase; C029063:fructose-1,6-diphosphate