Multiple forms of pyrophosphate:D-fructose-6-phosphate 1-phosphotransferase from wheat seedlings. Regulation by fructose 2,6-bisphosphate.
Two forms of pyrophosphate:D-fructose-6-phosphate 1-phosphotransferase have been isolated from wheat seedlings. One of these enzymes, termed PFP-1, has been purified to homogeneity. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicates that the enzyme is composed of two different polypeptide chains of Mr = 67,000 (alpha) and 60,000 (beta). PFP-1 has been assigned a molecular structure consisting of alpha 2 beta 2 based on an estimated Mr of 234,000 for the native enzyme. PFP-2, the other form of phosphotransferase, has also been purified extensively. Preliminary data suggest that the active form of PFP-2 is probably a dimer of a polypeptide chain of Mr = 60,000. Immunological studies indicate that the two enzyme preparations share common antigenic determinants. The two forms of enzyme have very similar kinetic properties. The phosphotransferases are activated by fructose 2,6-bisphosphate (Fru-2,6-P2) which lowers the Km of the enzymes for fructose 6-phosphate but not that for PPi. Interestingly, PFP-1 is significantly more active than PFP-2 in the absence of Fru-2,6-P2. Also, PFP-1 exhibits a greater affinity (Ka = 7 nM) than PFP-2 (Ka = 26 nM) for the activator. Based on kinetic, immunological, and physicochemical parameters, it is suggested that the two enzymic forms are related in that they share the same catalytic moiety, i.e. the 60,000-dalton or beta subunit. The beta subunit when in complex formation with the alpha subunit, as in PFP-1, becomes more active in the absence of Fru-2,6-P2 as well as exhibits a greater sensitivity toward the effector.
D004789:Enzyme Activation; D005635:Fructosediphosphates / Q000494:pharmacology*; D006598:Hexosediphosphates / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism*; D010944:Plants / Q000201:enzymology*; D014908:Triticum / Q000201:enzymology
D005635:Fructosediphosphates; D006598:Hexosediphosphates; D007527:Isoenzymes; C027652:fructose 2,6-diphosphate; D010770:Phosphotransferases; C022502:pyrophosphate-fructose 6-phosphate 1-phosphotransferase