Neutral amino acid transport in human synovial cells: substrate specificity of adaptative regulation and transinhibition.
Neutral amino acid transport was characterized in human synovial cells. The amino acids tested are transported by all three major neutral amino acid transport systems, that is, A, L, and ASC. The model amino acid 2-aminoisobutyric acid (AIB) was found to be a strong specific substrate for system A in synovial cells. When cells were starved of amino acids, the activity of AIB transport increased, reaching a maximum within 1 h. The stimulation of transport activity was not blocked by cycloheximide and would thus appear to be related to a release from transinhibition. Similarly, the decrease in the activity of AIB transport observed after the addition of alpha-methyl-aminoisobutyric acid (meAIB) appeared to be related to transinhibition. However, using a different approach, that is, amino acid starvation followed by incubation with 10 mM meAIB and transfer to an amino acid-free medium with or without cycloheximide supplementation, a clear increase in AIB uptake, due both to derepression and a release from transinhibition, was observed. Unlike human fibroblasts, the depression of system A in these synovial cells was not serum-dependent. The process of derepression was observed only after preloading with meAIB. Neither AIB nor alanine produced this phenomenon. Moreover, alanine preloading led to a large increase in AIB transport activity due to a release from transinhibition. These observations indicate that the process of derepression and release from transinhibition are specific to the substrates present in the culture medium prior to amino acid starvation.
Aussel
C
C
Laboratoire de Biochimie Générale, CNRS UA DO 622, Université Paris Sud, Chatenay-Malabry, France.
Rousseau-Loric
S
S
Cynober
L
L
Agneray
J
J
Ekindjian
O G
OG
D000409:Alanine / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism*; D000621:Aminoisobutyric Acids / Q000378:metabolism; D001692:Biological Transport; D005998:Glycine / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007930:Leucine / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D013583:Synovial Membrane / Q000378:metabolism*
D000596:Amino Acids; D000621:Aminoisobutyric Acids; C100049:2-aminoisobutyric acid; C017911:2-(methylamino)isobutyric acid; D008715:Methionine; D007930:Leucine; D000409:Alanine; D005998:Glycine