Kinetics of 6-phosphofructo-2-kinase from Saccharomyces cerevisiae: inhibition of the enzyme by ATP.
6-Phosphofructo-2-kinase (PFK-2) was purified from yeast and separated from fructose-2,6-biphosphatase (FBPase-2). The purification procedure involved polyethylene glycol fractionation followed by chromatography on DEAE-Sephacel. PFK-2 and FBPase-2 were copurified in these steps. Separation of the two enzymes resulted from Sephacryl S-300 Blue chromatography. Then, PFK-2 was chromatographed on CM-Sephadex and eluted with a gradient of KCl. Finally, PFK-2 was rechromatographed at CM-Sephadex and specifically eluted with fructose 6-phosphate. PFK-2 (specific activity 1.3 U/mg) was purified about 25,000-fold. The enzyme is inhibited by ATP which is particularly pronounced at low concentrations of magnesium and fructose 6-phosphate. Phosphoenolpyruvate and sn-glycerol 3-phosphate are inhibitors of the enzyme.
Bedri
A
A
Institute of Biochemistry, Karl-Marx University, Leipzig, GDR.
Kretschmer
M
M
Schellenberger
W
W
Hofmann
E
E
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D002850:Chromatography, Gel; D005994:Glycerophosphates / Q000494:pharmacology; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D010732:Phosphofructokinase-1 / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D005994:Glycerophosphates; D000244:Adenosine Diphosphate; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; D010732:Phosphofructokinase-1; D008274:Magnesium