Influence of age on 5'-nucleotidase in plasma membranes isolated from rat liver

Studies were carried out to determine the kinetic properties of 5'-nucleotidase from liver plasma membranes in rats of different ages; four groups were examined, namely rats 25 +/- 2, 60 +/- 5, 230 +/- 15 and 525 +/- 20 days old. The 5'-nucleotidase showed minimum Vmax values in young rats; differences in this kinetic parameter were not detected among the other groups. However, the Km values increased during growth and development, declined in young animals and increased again in the middle-aged. Arrhenius plots of the 5'-nucleotidase activity showed a single break at aroung 22 degrees C in developing the middle-aged animals; the break temperature decreased to 17 degrees C in the rats 230 +/- 15 days old. The pH-Vmax and pH-Km curves showed a maximum at pH 7.6 at all ages. Lipid analysis of membrane preparations was carried out. Phospholipid composition did not change markedly with age. The cholesterol level decreased between 25 +/- 2 and 60 +/- 5 days. The degree of saturation of fatty acids seemed to increase in the same period, but reached the lowest value in the young rats. The results indicate that either sphingomyelin or other phospholipids do not affect the isothermal kinetics of 5'-nucleotidase during development and aging. Furthermore, phospholipid polar groups as well as the cholesterol and fatty acids of the bulk lipid phase modulate the membrane fluidity in the same way at different ages. Finally, the modifications of the Km with age cannot be correlated with changes in the surface charge.