Characterization of carrier-mediated transport systems for small neutral amino acids in human fibroblast lysosomes.
Analog inhibition studies of the uptake of proline, serine, and threonine into human fibroblast lysosomes, purified on Percoll gradients, reveal the presence of three new transport systems. These systems fail to show the Na+ requirement usual for the plasma membrane. Proline uptake into fibroblast lysosomes occurs mainly by two routes: a predominant route half-saturating at 0.01 mM, and a lower-affinity route, half-saturating at 0.07 mM. The latter so far appears specific for L-proline and its 3,4-dehydro derivative. The high affinity route has a broad scope, recognizing best, beyond these two amino acids, various unbranched neutral amino acids not over 5 carbons long. Neither system accepts to a significant extent D-proline, hydroxyproline, cationic or anionic amino acids, nor neutral ones with bulky side chains. 2-Aminoisobutyrate and its N-methyl derivative have little effect on proline uptake, in contrast to their effectiveness on its uptake by the intact fibroblast. The rate of lysosomal proline uptake maximizes at about pH 6.4, is inversely related to the osmolarity of the medium, and is unaffected by the extralysosomal presence of MgATP. The competition among alanine, serine, and threonine points to sharing of the broad-scope system for proline, although the main part of their uptake occurs by a third route that rejects amino acids in which the alpha-amino group is methylated.
D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism*; D001693:Biological Transport, Active; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D007700:Kinetics; D008247:Lysosomes / Q000378:metabolism*; D008433:Mathematics; D009994:Osmolar Concentration; D011392:Proline / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D013912:Threonine / Q000378:metabolism
D000596:Amino Acids; D013912:Threonine; D012965:Sodium Chloride; D012694:Serine; D011392:Proline; D000409:Alanine