Alternative binding of two sequential glycolytic enzymes to microtubules. Molecular studies in the phosphofructokinase/aldolase/microtubule system.
Simultaneous binding of two sequential glycolytic enzymes, phosphofructokinase and aldolase, to a microtubular network was investigated. The binding of the phosphofructokinase to microtubules and its bundling activity has been previously characterized (Lehotzky, A., Telegdi, M., Liliom, K., and Ovádi, J. (1993) J. Biol. Chem. 268, 10888-10894). Aldolase binding to microtubules at near physiological ionic strength is weak (Kd = 20 microM) as compared with that of the kinase (Kd = 1 microM). The interactions of both enzymes with microtubules are modulated by their common intermediate, fructose-1,6-bisphosphate. Pelleting and electron microscopic measurements have revealed that the aldolase binding interferes with that of phosphofructokinase, although they have distinct binding domains on microtubules. The underlying molecular mechanism responsible for this finding is that in the solution phase aldolase and phosphofructokinase form a bienzyme complex that does not bind to the microtubule. The bienzyme complex formation does not influence the catalytic activity of aldolase, however, it inhibits the dissociation-induced inactivation of the kinase by stabilizing a catalytically active molecular form. The present data suggest the first experimental evidence that two sequential glycolytic enzymes do not associate simultaneously to microtubules, but their complexation in solution provides kinetic advantage for glycolysis.
Vértessy
B G
BG
Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, Budapest, H-1518, P.O.B. 7., Hungary.
Orosz
F
F
Kovács
J
J
Ovádi
J
J
D000818:Animals; D002384:Catalysis; D002417:Cattle; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D006019:Glycolysis; D007700:Kinetics; D008854:Microscopy, Electron; D008870:Microtubules / Q000378:metabolism*; D008954:Models, Biological; D009391:Nephelometry and Turbidimetry; D009994:Osmolar Concentration; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D011485:Protein Binding
D010732:Phosphofructokinase-1; D005634:Fructose-Bisphosphate Aldolase