Studies on the amino acid transport systems in the human placenta--L-alanine and L-leucine uptake by microvillous brush border membrane vesicles
Asai, M.
Nihon Sanka Fujinka Gakkai Zasshi 35(1): 10-18
1983
ISSN/ISBN: 0300-9165 PMID: 6298324 Document Number: 201478
To characterize the placental amino acid transport systems, L-alanine and L-leucine uptakes were studied using microvillous brush border membrane vesicles prepared from human placenta. The specific activities of alkaline phosphatase and 5'-nucleotidase in the membrane preparation were enriched 9-11 times as high as those in the homogenate. Intravesicular water (IVW) volume determined with 3-O-methyl-D-glucose was 0.59 microliters/mg protein. The saturation kinetics of L-leucine uptake by the vesicles equilibrated with Na+ gave a single set of Km (4.2mM) and Vmax (1.16 mumol/ml IVW/30s). These parameters were clearly different from those for L-alanine uptake reported previously (Asai et al.: Biochem. Int., 4:377, 1982). In the presence of an inward Na+-gradient L-leucine uptake was stimulated about 2 times, but transient accumulation was not observed differing from L-alanine uptake. Discrimination of the neutral amino acid transport systems in the presence of an inward 100mM Na+-gradient revealed that the relative contributions of A, ASC and L systems, and simple diffusion were 55, 20, 15 and 10% for L-alanine, and 45, 0, 15 and 40% for L-leucine, respectively. The results indicate that the neutral amino acid transport systems in the human placental microvillous membranes are clearly different between L-alanine and L-leucine.