Effect of pH and lysosomotropic agents on membrane-associated and internalized 125I-iodinated human growth hormone in cultured human lymphocytes: a quantitative biochemical and electron microscopic study.
When 125I-iodinated human GH ([125I]iodo-hGH) interacts with cultured human lymphocytes at 15 C, the reaction is reversible, but at 37 C the reaction becomes less dissociable as a function of incubation time. Acidification of the incubation medium results in rapid ligand dissociation at 15 C, but at 37 C the acid-dissociable component decreases as a function of incubation time. Under conditions where approximately 50% of the ligand is internalized by the cell, 90% is nondissociable. When the 37 C incubation is carried out in the presence of 25 mM NH4Cl, cell-associated radioactivity is increased. Under these conditions approximately 90% of cell-associated radioactivity also is nondissociable. Using quantitative electron microscopic autoradiography, the proportion of [125I]iodo-hGH associated with the plasma membrane and internalized by the cell is indistinguishable in the presence or absence of NH4Cl. Irreversible [125I]iodo-hGH association with cultured human lymphocytes is due to time- and temperature-dependent effects in the plasma membrane of the cell. These effects cannot be distinguished from internalization by acidification. Furthermore, lysosomotropic agents increase cell-associated radioactivity, but the proportion internalized is not increased.
D000643:Ammonium Chloride / Q000494:pharmacology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D013006:Growth Hormone / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007457:Iodine Radioisotopes; D007700:Kinetics; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D008854:Microscopy, Electron; D013696:Temperature
D007457:Iodine Radioisotopes; D000643:Ammonium Chloride; D013006:Growth Hormone