Characterization of lysosomal monoiodotyrosine transport in rat thyroid cells. Evidence for transport by system h.
Lysosomal transport of monoiodotyrosine was characterized in countertransport experiments using rat FRTL-5 thyroid cell lysosomes. Monoiodotyrosine carrier activity was temperature-dependent (Ea = 11.65 kcal/mol) and had a pH optimum of 7.5. Carrier activity was minimally inhibited by KCl and NaCl, but unaffected by the presence of other ions or ATP. Monoiodotyrosine transport was unaffected by the presence of carbonyl cyanide m-chlorophenylhydrazone, nigericin, or ammonium chloride, indicating that a proton or K+ gradient is not necessary for monoiodotyrosine transport across the lysosomal membrane. Monoiodotyrosine countertransport showed a 6-fold increase in lysosomes from FRTL-5 cells grown in medium containing thyrotropin by comparison to cells grown without this hormone. Thyrotropin responsiveness raised the possibility that monoiodotyrosine was transported by system h, the only known lysosomal carrier whose activity is enhanced by thyrotropin. Consistent with this, monoiodotyrosine-loaded lysosomes exhibited countertransport of [3H]tyrosine, [3H]phenylalanine, and [3H]leucine, three system h ligands, but not [3H]cystine, a nonsystem h ligand. Unlabeled tyrosine, phenylalanine, and leucine, but not cystine or proline, inhibited [125I]monoiodotyrosine countertransport, and leucine inhibition of [3H]tyrosine countertransport and [125I]monoiodotyrosine countertransport yielded virtually identical KI values, 3.5 and 3.2 microM, respectively. Competition studies with monoiodotyrosine analogues showed that system h recognizes a broad range of ligands with an alpha-amino acid configuration at one end and a hydrophobic region at the other. Ring-substituted halogens, regardless of mass or ring position, but not amino, nitro, hydroxy, or methoxy groups, enhanced carrier recognition of system h analogues. It appears that a single system effects the transport of iodinated (e.g. monoiodotyrosine) and noniodinated (e.g. tyrosine) thyroglobulin catabolites into the cytosol for salvage and reutilization by FRTL-5 thyroid cells.
Andersson
H C
HC
Section on Human Biochemical Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Kohn
L D
LD
Bernardini
I
I
Blom
H J
HJ
Tietze
F
F
Gahl
W A
WA
D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism*; D007470:Monoiodotyrosine / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D051381:Rats; D013329:Structure-Activity Relationship; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D013972:Thyrotropin / Q000494:pharmacology; D014443:Tyrosine / Q000378:metabolism
D002352:Carrier Proteins; D014443:Tyrosine; D010649:Phenylalanine; D013972:Thyrotropin; D007470:Monoiodotyrosine; D007930:Leucine