A new transient activator of phosphofructokinase during initiation of rapid glycolysis in brain.
The tissue contents of previously known allosteric effectors of brain phosphofructokinase (EC 2.7.1.11) (PFK) and the kinetic behavior of isolated PFK were investigated during the initiation of rapid glycolytic flux in freeze-blown rat brain. Comparing 0- with 5-s brains revealed that there was a 4-fold drop in total tissue content of Fru-6-P and a 5.6-fold increase in Fru-1,6-P2 consistent with activation of PFK. Additionally, analysis of brain content showed a 15-fold increase in AMP, a 3-fold decrease in ATP, a 3-fold decrease in Pi, and a 1.6-fold increase in NH4+. There was no change in Fru-2,6-P2, H+, citrate, or Glc-1,6-P2 or the kinetic profiles of isolated PFK for ATP inhibition or Fru-2,6-P2 activation. We concluded that the observed change in PFK activity could be accounted for only partially by changes in the concentrations of adenine nucleotides and other known effectors. High performance liquid chromatography fractions of extracts obtained from 5-s brains showed the activator with a mobility identical to ribose 1,5-P2 and gave 2 nmol/g (wet weight) at 0 s, 10 nmol/g at 5 s, and 2 nmol/g at 20 s. Assay of PFK in the presence of effectors determined to be in tissue at 5 s showed that addition of 10 nmol/ml ribose 1,5-P2 gave a 4-fold activation of PFK. Based on the rapidity of its formation, its potency of activation, and its similarity in chemical properties to authentic ribose 1,5-P2, we conclude that ribose 1,5-P2 served as the initial activator of PFK in brain.
Ogushi
S
S
Department of Veterans Affairs Medical Center, Dallas, Texas 75216.
Lawson
J W
JW
Dobson
G P
GP
Veech
R L
RL
Uyeda
K
K
D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation / Q000187:drug effects; D005635:Fructosediphosphates / Q000378:metabolism; D005636:Fructosephosphates / Q000378:metabolism; D006019:Glycolysis; D007700:Kinetics; D008297:Male; D010428:Pentosephosphates / Q000378:metabolism / Q000494:pharmacology*; D010710:Phosphates / Q000378:metabolism; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D005635:Fructosediphosphates; D005636:Fructosephosphates; D010428:Pentosephosphates; D010710:Phosphates; D000644:Quaternary Ammonium Compounds; C050562:ribose-1,5-bisphosphate; D000249:Adenosine Monophosphate; C027618:fructose-6-phosphate; D000255:Adenosine Triphosphate; D010732:Phosphofructokinase-1; C029063:fructose-1,6-diphosphate