General properties of 14C-L-Valine-binding to human brain tissue

L-Valine (VAL) binding is saturable, reversible, linear within a protein concentration of 0.2 to 0.8 mg prot./ml, pH dependent (pH-optimum 7.1 to 7.4), temperature dependent with extremely low binding at 0-4 degrees C; the assay runs at 37 degrees C; it is ion dependent (Bmax:CaCl2 (5mM): + 100%; NaCl (5 mM): -50%); very sensitive to TRITON X-100 (Bmax at 0.01%; -70%); N-ethylmaleimide (0.5 mM): Competitive inhibition; no change in non-specific binding; there are no effects by puromycin plus GTP; VAL binds to synaptic membranes of human frontal cortex with KD-values several fold lower than that observed with homogenates, the distribution of VAL-binding is different in various human brain areas and can be inhibited by L-leucine, L-isoleucine, L-serine, DL-threonine, glycine and D-valine, while aromatic amino acids, beta-alanine, GABA, taurine, glutamate, proline and gamma-glutamyltaurine have no effects of 10(-3) M.