3'-Azido-3'-deoxythymidine reduces the rate of transferrin receptor endocytosis in K562 cells.
K562 cells, exposed for at least 24 h to 5 microM 3'-azido-3'-deoxythymidine (AZT), gave rise to an overall increase in the number of cell surface transferrin binding receptors (18-20%). This effect was ascertained either with binding experiments by using 125I-transferrin and with immunoprecipitation by using a specific monoclonal antibody against the transferrin receptor. At higher AZT concentrations (20 and 40 microM), a further increase was found, that is, up to 23% by binding experiments and up to 110% by immunoprecipitation. However, Scatchard analysis of the binding data indicated that although the number of cell surface transferrin receptors increased, the affinity of transferrin for its receptor did not change (Ka=4.0x108 M). Surprisingly, immunoprecipitation of total receptor molecules showed that the synthesis of receptor was not enhanced by the drug treatment. The effect of AZT on transferrin internalization and receptor recycling was also investigated. In this case, data indicated that the increase in the number of receptors at the cell surface was probably due to a slowing down of endocytosis rate rather than to an increased recycling rate of the receptor to cell surface. In fact, the time during which half the saturated amount of transferrin had been endocytosed (t1/2) was 2.15 min for control cells and 3.41, 3.04, and 3.74 min for 5, 20, and 40 microM AZT-treated cells, respectively. Conversely, recycling experiments did not show any significant differences between control and treated cells. A likely mechanism through which AZT could interfere with the transferrin receptor trafficking, together with the relevance of our findings, is extensively discussed.
Department of Biomedical Sciences and Technology, University of L'Aquila, Via Vetoio Coppito, I-67100, L'Aquila, Italy.
A M
AM
D'Alessandro
G
G
D'Andrea
L
L
Di Ciccio
F
F
Brisdelli
A C
AC
Rinaldi
A
A
Bozzi
A
A
Oratore
D002250:Carbon Radioisotopes; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D007501:Iron / Q000378:metabolism; D007504:Iron Radioisotopes; D020014:K562 Cells; D011233:Precipitin Tests; D011485:Protein Binding; D011990:Receptors, Transferrin / Q000187:drug effects* / Q000378:metabolism; D015215:Zidovudine / Q000494:pharmacology*
D002250:Carbon Radioisotopes; D007504:Iron Radioisotopes; D011990:Receptors, Transferrin; D015215:Zidovudine; D007501:Iron