title: Direct demonstration of rapid insulin-like growth factor II Receptor internalization and recycling in rat adipocytes. Insulin stimulates 125I-insulin-like growth factor II degradation by modulating the IGF-II receptor recycling process.
abstract: The photoactive insulin-like growth factor (IGF)-II analogue 4-azidobenzoyl-125I-IGF-II was synthesized and used to label specifically and covalently the Mr = 250,000 Type II IGF receptor. When rat adipocytes are irradiated after a 10-min incubation with 4-azidobenzoyl-125I-IGF-II at 10 degrees C and immediately homogenized, most of the labeled IGF-II receptors are associated with the plasma membrane fraction, indicating that receptors accessible to the labeling reagent at low temperature are on the cell surface. However, when the photolabeled cells are incubated at 37 degrees C for various times before homogenization, labeled IGF-II receptors are rapidly internalized with a half-time of 3.5 min as evidenced by a loss from the plasma membrane fraction and a concomitant appearance in the low density microsome fraction. The low density microsomes were previously shown to contain intracellular membranes (Oka, Y., and Czech, M.P. (1984) J. Biol. Chem. 259, 8125-8133). The steady state level of cell surface IGF-II receptors in the presence or absence of IGF-II, measured by the binding of anti-IGF-II receptor antibody to cells, remains constant under these conditions, demonstrating that IGF-II receptors rapidly recycle back to the cell surface at the same rate as receptor internalization. Using the above methodology, it is shown that acute insulin action: 1) increases the steady state number of cell surface IGF-II receptors; 2) increases the number of ligand-bound IGF-II receptors that are internalized per unit of time, as evidenced by a large increase in the photolabeling of intracellular membrane IGF-II receptors when cells are incubated at 37 degrees C with insulin and 4-azidobenzoyl-125I-IGF-II prior to photoactivation; and 3) increases the rate of cellular 125I-IGF-II degradation by a process that is blocked by anti-IGF-II receptor antibody. The results indicate that the action of insulin to elevate the steady state number of cell surface IGF-II receptors leads to an increased internalization flux of IGF-II-bound receptors, mediating increased IGF-II uptake and degradation.
authors.forename: Y
authors.initials: Y
authors.lastname: Oka
authors.forename: L M
authors.initials: LM
authors.lastname: Rozek
authors.forename: M P
authors.initials: MP
authors.lastname: Czech
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism*; D000345:Affinity Labels / Q000138:chemical synthesis; D000818:Animals; D001386:Azides / Q000138:chemical synthesis; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology*; D007335:Insulin-Like Growth Factor II / Q000031:analogs & derivatives*; D007457:Iodine Radioisotopes; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism*; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000138:chemical synthesis / Q000378:metabolism*
chemical_list: C046102:4-azidobenzoyliodo-insulin-like growth factor II; D000345:Affinity Labels; D001386:Azides; D007328:Insulin; D007457:Iodine Radioisotopes; D010455:Peptides; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D013002:Somatomedins; D007335:Insulin-Like Growth Factor II