High-affinity interleukin 2 receptor alpha and beta chains are internalized and remain associated inside the cells after interleukin 2 endocytosis.
High-affinity interleukin 2 (IL2) receptors on human T lymphocytes are multimeric complexes containing two IL2-binding polypeptides, alpha and beta chains of 50-55 and 70-75 kDa, respectively, associated by noncovalent bonds. IL2 binds to high-affinity IL2 receptors on the surface of T lymphocytes, mediates cell growth, and is internalized. In this paper, we used a biochemical method to directly identify the receptors components internalized together with the ligand. 125I-IL2-receptor complexes were solubilized with the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate, and IL2-binding polypeptides were identified by cross-linking with disuccinimidyl suberate. Under such conditions, the noncovalent association between alpha and beta is maintained. After IL2 internalization, two complexes of about 70 and 90 kDa, IL2 crosslinked to alpha and beta, respectively, were found inside the cells. Both components were immunoprecipitated with either anti-alpha or anti-beta monoclonal antibodies. This shows that the alpha and beta chains are found in an intracellular compartment after IL2 endocytosis, and remain associated as a ternary complex with IL2.
Duprez
V
V
Unité de Biologie des Interactions Cellulaires, Centre National de la Recherche Scientifique URA 361, Institut Pasteur, Paris, France.
Ferrer
M
M
Dautry-Varsat
A
A
D002793:Cholic Acids; D003432:Cross-Linking Reagents; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007700:Kinetics; D015375:Receptors, Interleukin-2 / Q000378:metabolism*; D013388:Succinimides / Q000737:chemistry; D014407:Tumor Cells, Cultured
D002793:Cholic Acids; D003432:Cross-Linking Reagents; D003902:Detergents; D007376:Interleukin-2; D007457:Iodine Radioisotopes; D015375:Receptors, Interleukin-2; D013388:Succinimides; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; C019358:disuccinimidyl suberate