Transport of L-histidine by human diploid fibroblasts in culture

Abstract

The transport of L-histidine has been characterized in skin derived diploid human fibroblasts, cultured under strictly controlled conditions. The transport measurements were made on cells grown to subconfluency after 60 to 90 min timed preincubation. The data, at substrate concentrations ranging from 0.050 to 10 mmol/l, were analyzed by a computer program. A saturable transport system (Km = 0.25 mmol/l, Vmax = 17 nmol/mg protein per min) and a nonsaturable component of influx (Kd = 1.6 +/- 0.4 nmol/mg protein/min per mmol) were found. L-Histidine displayed no Na+ requirement at either low or high concentrations. Inhibition analysis demonstrated that L-histidine uptake at low concentration was poorly inhibited by amino acids known to be effective inhibitors of system A. The largest fraction of L-histidine uptake was inhibited by 2-amino-bicyclo (2,2,1)-heptane-2-carboxylic acid (BCH), leucine, and tryptophan. These results indicated that L-histidine is transported in human fibroblasts mainly by the Na+ independent system L. The differences between this cell type and others studied previously are discussed.