potassium-thiocyanate and fructose-2-6-diphosphate

potassium-thiocyanate has been researched along with fructose-2-6-diphosphate* in 1 studies

Other Studies

1 other study(ies) available for potassium-thiocyanate and fructose-2-6-diphosphate

ArticleYear
Pyrophosphate-dependent phosphofructokinase from the amoeba Naegleria fowleri, an AMP-sensitive enzyme.
    The Biochemical journal, 1993, Jun-15, Volume: 292 ( Pt 3)

    PPi-dependent phosphofructokinase (PPi-PFK) was detected in extracts of the amoeba Naegleria fowleri, with a specific activity of about 15-30 nmol/min per mg of protein, which was increased about 2-fold by 0.5 mM AMP. PPi-PFK was inactivated upon gel filtration and could be re-activated by incubation at 30 degrees C in the presence of AMP. N. fowleri PPi-PFK was purified more than 1100-fold to near homogeneity with a yield of about 25%. The pure enzyme had a specific activity of 65 mumol/min per mg of protein, and SDS/PAGE analysis showed a single band, of 51 kDa. Size-exclusion chromatography revealed the existence of two forms: a large one (approximately 180 kDa), presumably a tetramer, which was active, and a smaller one (approximately 45 kDa), presumably the monomer, which was inactive, but could be re-activated and converted into the large form by incubation at 30 degrees C in the presence of 0.5 mM AMP. Reactivation was also observed at 30 degrees C in the absence of AMP, particularly at higher enzyme concentration or in the presence of poly(ethylene glycol). Inactivation of the tetrameric enzyme was promoted by 0.25 M potassium thiocyanate. The enzyme displayed Km values of 10 and 15 microM for fructose 6-phosphate and PPi, respectively, in the forward reaction, and of 35 and 590 microM for fructose 1,6-bisphosphate and Pi in the backward reaction. The activity was dependent on the presence of Mg2+. AMP increased Vmax. about 2-fold without changing the affinity for the substrates; its half-maximal effect was observed at 2 microM.

    Topics: Adenosine Monophosphate; Animals; Chromatography, Gel; Chromatography, Ion Exchange; Detergents; Diphosphates; Electrophoresis, Polyacrylamide Gel; Enzyme Activation; Fructosediphosphates; Kinetics; Macromolecular Substances; Molecular Weight; Naegleria fowleri; Octoxynol; Phosphofructokinase-1; Polyethylene Glycols; Spectrophotometry, Ultraviolet; Substrate Specificity; Thiocyanates

1993