Single-site uptake of neutral amino acids into guinea-pig intestinal rings

1. A kinetic analysis of amino acid influx into guinea-pig small intestinal rings has been performed in an attempt to ascertain whether one or more transport sites for these substrates exists in the luminal membrane of the enterocyte. 2. No indirect correction for uptake into the extracellular space was applied, but it was assumed in the analysis that the total uptake included a diffusion term. This procedure was vindicated by the results obtained. 3. All analyses were performed by non-linear regression techniques. In many experiments, both substrate and inhibitor concentrations were varied within the same experiment, thus giving rise to three-dimensional diagrams describing transport processes. 4. All results pointed to the existence of a single transport agency shared by all amino acids tested. The kinetic constants, Km and Vmax, were independent of the concentration range used for their estimation; such behaviour would not be expected if several sites with different kinetic constants were available for transport. The value of KD, the constant describing the diffusive component of uptake, was the same when estimated from the uptake of an amino acid alone or when determined as the asymptote of the curve describing the inhibition of this uptake by an analogue. Finally, the Ki for an amino acid when used as an inhibitor was identical to its Km when used as substrate. This property was maintained even when the most disparate pair of amino acids, threonine and isoleucine, was examined.