A comparison of the rate equations, kinetic parameters, and activation energies for the initial uptake of L-lysine, L-valine, gamma-aminobutyric acid, and alpha-aminoisobutyric acid by mouse brain slices

Abstract

At substrate concentrations, in medium, of 0.2 to 20 mM and at temperatures of 25 and 37 degrees C, the initial concentrative influx of the amino acids L-lysine (30 and 37 degrees C), L-valine, and gamma-aminobutyric acid into incubated mouse-cerebrum slices follows the rate equation for the initial influx of alpha-aminoisobutyric acid (Cohen, J. Physiol. 228:105, 1973), v equals Vmax/(1+Kt/S)+kuS. Kinetic constants at 37 degrees C are: Vmax equals 0.089 mumoles/g final wet wt of slices, min, Kt equals 0.69 mM, ku equals 0.037 mumoles/g final wet wt, mM-substrate, min for L-lysine; Vmax equals 0.60, Kt equals 1.30, ku equals 0.067 for L-valine; and Vmax equals 1.71, Kt equals 1.58, ku equals 0.094 for gamma-aminobutyric acid. The linear term, kuS, is due to an unsaturable process of concentrative uptake, not diffusion. Comparison of temperature coefficients reveals a "reference" pattern for typical low affinity transport of amino acids into brain slices. Its characteristics are: Activation energies associated with Vmax and ku are in range 14 to 20 kcal/mole; K, varies only slightly with temperature, L-Lysine and alpha-aminoisobutyric acid fit this pattern; L-valine and gamma-aminobutyric acid deviate in part. The Akedo-Christensen plot (J. Biol. Chem. 237:118, 1962) does not distinguish between the rateequation v equals Vmax/(1+Kt/S)+kuS for saturable uptake plus first-order unsaturable concentrative uptake, and the rate equation v equals Vmax/(1 + Kt/S)+kd(S minus Si) for saturable uptake plus first-order nonconcentrative "passive diffusion".