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Query: UNIPROT:P02794 (ferritin)
 17,525 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We previously reported that in 3T3-L1 adipocytes 125I-insulin associates preferentially with microvilli and coated pits at low temperatures and early times of incubation. At higher temperatures it is internalized through a series of membrane limited intracellular compartments. In the present study, we used a high resolution probe, cationic ferritin (CF), to track adsorptive endocytosis in the 3T3-L1 adipocyte. We find that CF initially associates with coated pits at 2 min of incubation at 37 degrees C. With further incubation at 37 degrees C CF is internalized and after 2 to 10 min of incubation is predominantly localized to coated and non-coated clear vesicles. Approximately 50% of the apparent coated vesicles seen near the plasma membrane on single thin sections are shown by serial sectioning to be true vesicles (i.e., without a surface connection). At later time points CF is localized predominantly to lysosomal structures and, to a much smaller extent, Golgi-related structures. The remarkable similarity between 125I-insulin and CF with respect to post-binding processing suggests that while the membrane receptor confers the initial specificity, post-binding events are common for different types of ligands after they bind to cell surfaces and are subject to adsorptive endocytosis.
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PMID:Surface interactions and intracellular localization of cationic ferritin: similarity to 125I-insulin in 3T3-L1 adipocytes. 613 62

Ruthenium red binding demonstrates that the extensive microvesicle system of isolated rat adipocytes is, for the most part, open to and continuous with the plasmalemma proper. Morphometric estimates indicate that insulin treatment has no effect on the relationship between microvesicles and the cell surface. Neither does insulin affect the apparent lack of pinocytotic activity of these vesicles as judged by a time course analysis of cells incubated with horseradish peroxidase, which is bound to the membrane of the vesicles, but is not internalized. Insulin does produce a small but repeatable and measurable increase in average diameter of the microvesicles from 73 to 78 nm. Unlike the positively charged ruthenium red, which binds to both plasmalemmal as well as microvesicular surfaces, cationic ferritin did not readily bind to microvesicle membranes, a result indicating some distinction between the two membrane surfaces. The implications of the lack of dramatic visible morphological effects of insulin upon the adipocyte plasmalemma and it's associated microvesicles are discussed in light of the proposed role of insulin as a mediater of translocation of membrane-associated transporters to and from the cytoplasm.
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PMID:The relationship of microvesicles to the plasmalemma of rat adipocytes. 619 5

Previous studies have shown that serum levels of the somatomedin, nonsuppressible insulin-like activity (NSILA-S), are extremely low in patients with thalassemia major. Since these patients are not GH deficient, several other possible mechanisms for the reduced levels of NSIL-S have been explored. No evidence for the presence of NSILA inhibitors was obtained either in mixing experiments of normal serum and thalassemic sera or after acid gel chromatography of thalassemic sera. The high iron and ferritin levels of thalassemia had no effects on the NSILA-S bioassay itself or on the binding of GH to its hepatic receptors. GH molecules secreted as a result of exercise-induced GH stimulation tests were shown to be both immunologically and biologically reactive. No circulating GH-binding proteins were present in thalassemic sera. Since the liver function in the group of patients included in this study was only slightly abnormal, it is considered unlikely that generalized hepatic damage due to the severe iron overload of thalassemia is a major cause. These results suggest that neither NSILA-S inhibitors, abnormal GH molecules, nor hepatic damage contribute to the failure of these patients to produce NSILA-S and that a specific defect may exist at the hepatic GH receptor or postreceptor level.
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PMID:Studies on the possible mechanism for deficiency of nonsuppressible insulin-like activity in thalassemia major. 625 46

In this ultrastructural study, monomeric ferritin-insulin was used to further elucidate the role of disulfide bonds in maintaining the natural groups of insulin receptors on adipocyte plasma membranes. Dithiothreitol (1 mM) caused partial disruption of the occupied receptor groups with an increase in single receptors to greater than 50% of total occupied receptors. N-Ethylmaleimide (1 mM) disrupted the groups to the same extent as dithiothreitol and the effect was partly additive with the dithiothreitol effect. The magnitude of the disruption caused by dithiothreitol or N-ethylmaleimide was similar to that caused by cytochalasin B. Dithiothreitol, a reducing agent, caused a marked increase in binding of insulin to the plasma membranes while N-ethylmaleimide and cytochalasin B, both thiol reagents, had little if any effect on insulin binding. These data suggest that two different sets of disulfide bonds are involved. One set was susceptible to both reducing and thiol reagents and responsible for holding the receptor groups together, and the other set was susceptible to reducing agents only and related to the increased insulin binding caused by dithiothreitol. A proposed model is discussed.
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PMID:Partial disruption of naturally occurring groups of insulin receptors on adipocyte plasma membranes by dithiothreitol and N-ethylmaleimide: the role of disulfide bonds. 634 87

Surface labeling and internalization of insulin was demonstrated ultrastructurally with human peripheral lymphocytes and with "activated"/transformed lymphocytes from mitogen-treated cultures using the colloidal gold-labeled insulin-bovine serum albumin (GIA) procedure. The majority of peripheral lymphocytes bound only limited amounts of the insulin complex, while approximately 15% of the lymphocyte population bound modest to comparatively large quantities of the labeled hormone. Quantitative labeling data indicated a skewed GIA labeling continuum for peripheral lymphocytes rather than separate, distinct populations. Sequential labeling studies with the GIA complex followed by either the ferritin-conjugated goat anti-human immunoglobulin or the E-rosette techniques indicated that insulin labeling was neither T nor B cell specific, since extremes of GIA labeling were found in both populations. Many, but not all, circulating lymphocytes with elevated insulin binding had morphological features suggestive of "active" cells, viz., larger cell, nuclear, nucleolar, and Golgi sizes, dispersed chromatin, and greater numbers of polysomes than lymphocytes having minimal GIA labeling. Both phytohemagglutinin (PHA), a T-cell mitogen, and pokeweed mitogen (PWM), a B/T cell mitogen, induced an increase in mean GIA labeling of cultured lymphoid cells as compared to non-mitogen-treated controls. The majority of mitogen-transformed "blast-like" cells had more extensive insulin labeling than nontransformed small (medium)-size lymphocytes, although an overlap in labeling densities was noted in these two groups. PHA induced a slight increase in mean surface GIA labeling of the nontransformed lymphocyte population at 48 and 72 hr of culture as compared to similar cells in non-mitogen-treated controls and PWM cultures. We interpret these findings as indicating the emergence of increased numbers of insulin binding sites on lymphocytes, both those in the circulation and in mitogen-treated cultures, during the early response (activation) to functional and/or metabolic modulations of the cell; this surface change does not appear to be directly related to blastogenic transformation.
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PMID:Insulin complex binding to human peripheral and mitogen-stimulated lymphocytes. 637 Nov 32

Monomeric ferritin-insulin was used as an ultrastructural marker to determine by quantitative electron microscopy the time course and route of insulin uptake in rat adipocytes. To approximate steady state membrane binding conditions prior to any internalization, adipocytes were prefixed with glutaraldehyde and incubated for 30 min with 70 nM monomeric ferritin-insulin. Electron micrographs of these cells showed that the ferritin-insulin particles were predominantly in small groups of receptor sites on the plasma membrane and in pinocytotic-like invaginations of the plasma membrane. Significant amounts of ferritin-insulin were observed in cytoplasmic vesicles of unfixed cells as early as 2 min and in multivesicular bodies and lysosome-like structures within 5 to 10 min after the addition of the ligand. Ferritin-insulin accumulation reached steady state levels in the cytoplasmic vesicles in 5 to 10 min and in the lysosome-like structures in 15 min. Little ferritin-insulin was bound to coated pits, and the relative paucity of coated pits found in adipocytes suggested that these specialized endocytotic structures have a relatively insignificant role in insulin uptake in fat cells. Quantitative analysis of the uptake process suggested that a proportion of the insulin internalized by the cell may not be transported to lysosomes, but may be recycled along with the insulin receptor to the plasma membrane.
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PMID:Quantitative ultrastructural analysis of receptor-mediated insulin uptake into adipocytes. 640 13

A quantitative morphological analysis of insulin uptake into adipocytes was undertaken to determine the structural basis for chloroquine-induced increases in intracellular insulin. Adipocytes were incubated with ferritin-labeled insulin in the presence or absence of 50 microM chloroquine at 37 degrees C for 2-90 min and the uptake of the hormone conjugate was determined quantitatively. Quantitative morphometry of cellular organelles also was performed. Chloroquine treatment of adipocytes incubated with 70 nM ferritin-labeled insulin resulted in: (i) a 120% increase in the number of lysosomes in the cytoplasm; (ii) a 75% increase in the average concentration of ferritin-labeled insulin in a lysosome; and (iii) a 25% increase in the percentage of lysosomes containing ferritin-labeled insulin. The cumulative result of these effects was a substantial increase in the amount of intact intracellular hormone within the lysosomes. These morphological data are consistent with biochemical data concerning chloroquine-induced accumulation of 125I-labeled insulin in adipocytes.
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PMID:Ultrastructural basis for chloroquine-induced increase in intracellular insulin in adipocytes: alteration of lysosomal function. 676 Jan 94

When 125I-labeled insulin (125I-insulin) is incubated with 3T3-L1 adipocytes and cells processed for electron microscopic autoradiography, the ligand initially localizes preferentially to microvilli and coated pits. As a function of time and temperature, this initial preferential localization to microvilli is lost, and the ligand is internalized by the cell. Serial sections of apparent coated vesicles near the cell surface indicate that about half of these structures are true vesicles and, therefore, intermediates in this receptor-mediated endocytotic process. With time, 125I-insulin localizes to larger intracellular membrane-bounded structures. When cells are incubated with another ligand, cationic ferritin, that is taken up by adsorptive endocytosis, essentially the same structures are involved as for the endocytosis of 125I-insulin. The data suggest that specificity for receptor-mediated endocytosis is conferred by the specific ligand receptor and possibly by ligand-induced receptor mobility in the plane of the plasma membrane. Other structures such as coated pits, coated vesicles, larger vesicles, and secondary lysosomes are common for different ligands.
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PMID:Receptor-mediated endocytosis of insulin: role of microvilli, coated pits, and coated vesicles. 681 48

We have purified a protein (Mr approximately 71,000) from murine sera 104-fold which directly binds biologically active phorbol esters, ingenol esters, and mezerein in a specific, reversible, and saturable manner. The binding of labeled phorbol-12,13-dibutyrate (PDBu) to the purified protein is rapid and dose-dependent. Those phorbol and ingenol esters which stimulate cell growth in culture and have tumor-promoting activity in vivo inhibit the binding of labeled PDBu, while the biologically inactive derivatives fail to do so. Other nonditerpene tumor promoters, retinoids, steroids, and prostaglandins do not interfere with PDBu-protein interaction. Epidermal growth factor, insulin, bovine serum albumin, hemoglobin, ovalbumin, ferritin, myoglobin, fetuin, and lipase do not interact directly with PDBu. The purified binding protein competitively inhibits the binding of PDBu to its specific receptors. It is nonglycosylated and slightly hydrophobic. The protein is heat- and acid-labile and is present in sera of various mammalian species. Its concentration in murine sera is age-, sex-, and strain-independent.
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PMID:Partial purification and characterization of a binding protein for biologically active phorbol and ingenol esters from murine sera. 694 92

In this study we used chloroquine to characterize the interalization and lysosomal degradation of receptor-bound 125I-insulin by rat adipocytes and to determine the role of lysosomal processing of insulin in the short-term biologic effects of the hormone. Chloroquine inhibited the degradation of 125I-insulin bound to adipocytes by both association and disslociation experiments. In the former experiments, chloroquine caused a time- and concentration-dependent increase in specifically bound insulin owing to an increase in intact insulin and a decrease in degradation products, as determined by trichloroacetic acid precipitability and gel chromatographic analysis of material extracted from the cells. In the dissociation experiments, 50 microM chloroquine decreased the rate of degradation by two third, as reflected in the release of degradation to or degraded by isolated plasma membranes, on the degradation of 125I-insulin by proteases in the incubation medium, or on the endocytotic uptake of receptor-bound insulin. Quantitative electron miroscopy, using monomeric ferritin-insulin, showed 50 microM chloroquine doubled the number of lysosomal structures containing ferritin. These findings are consistent with an inhibition by chloroquine of lysosomal degradation of internalized receptor-bound insulin. Chloroquine, at these same concentrations, had no effect on the ability of insulin to stimulate glucose transport and oxidation or to inhibit epinephrine-stimulated lipolysis. In these studies, we show that lysosomal degradation of internalized receptor-bound insulin is not necessary for insulin to cause short-term biologic effects in the adipocyte.
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PMID:Lysosomal degradation of receptor-bound 125I-labeled insulin by rat adipocytes: its characterization and dissociation from the short-term biologic effects of insulin. 699 35


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