Alteration of enzymes in ageing human fibroblasts in culture. II. Conditions for the reversibility and the mechanism of the alteration of glucose 6-phosphate dehydrogenase.
The appearance of a heat-labile glucose 6-phosphate dehydrogenase fraction can be induced by incubation of human fibroblasts at 4 degrees C and pH 7.4. Using this system we first studied the effects of pH and NADP+ on the reversibility of the alteration. In young cells, the induced alteration of glucose 6-phosphate dehydrogenase disappeared if the pH of the medium was lowered to 6.5 or if NADP+ was added. In old cells, the disappearance of the natural heat-labile glucose 6-phosphate dehydrogenase was possible only if both pH 6.5 and NADP+ were present. Secondly, we studied the effects of the alteration and its reversibility on the equilibrium between the enzyme subunits; we showed that alteration and inactivation are linked to a dissociation of the enzyme subunits into inactive monomers. We also proposed a model where the heat-labile enzyme is a transient form between the active dimer and inactive monomer. Our results are in perfect agreement with the theories of post-translational modifications now proposed to explain the presence of the altered enzymes in old cells.
D002470:Cell Survival; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000201:enzymology*; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism*; D009249:NADP / Q000378:metabolism
D007527:Isoenzymes; D009249:NADP; D005954:Glucosephosphate Dehydrogenase