scholarly journals The stimulus-sensitive H2O2-generating system present in human fat-cell plasma membranes is multireceptor-linked and under antagonistic control by hormones and cytokines

1995 ◽  
Vol 307 (2) ◽  
pp. 543-548 ◽  
Author(s):  
H I Krieger-Brauer ◽  
H Kather
Keyword(s):  
Growth Factor ◽  
Plasma Membranes ◽  
Fat Cells ◽  
Inhibitory Effects ◽  
Fat Cell ◽  
Redox Equilibrium ◽  
Necrosis Factor Alpha ◽  
H2o2 Generation ◽  
Generating System ◽  
Antagonistic Control

Previous work demonstrated that human fat-cells possess a plasma-membrane-bound H2O2-generating system that is activated by insulin. Here we show that this system is under antagonistic control by various hormones and cytokines that typically act through several distinct receptor families. Similarly to insulin, oxytocin and tumour necrosis factor alpha acted as stimulators of NADPH-dependent H2O2 generation, whereas isoprenaline, a beta-adrenergic agonist, had inhibitory effects. Surprisingly, the acidic and basic isoforms of fibroblast growth factor as well as homodimeric platelet-derived growth factor AA and BB had antagonistic stimulatory and inhibitory effects on NADPH-dependent H2O2 generation. The agents tested acted at discrete ligand-specific receptors and their mechanisms of action were membrane-delimited and occurred in the absence of ATP. These findings implied that established pathways of signal transduction, including receptor kinases or second-messenger-dependent protein kinases A and C, were not involved and placed the stimulus-sensitive H2O2-generating system in a position comparable with adenylate cyclase. It was concluded that the stimulus-sensitive H2O2-generating system of human fat-cells meets all criteria of a universal signal-transducing system for hormones and cytokines that may link ligand binding to cell-surface receptors to changes in the intracellular redox equilibrium.

scholarly journals Antagonistic effects of different members of the fibroblast and platelet-derived growth factor families on adipose conversion and NADPH-dependent H2O2 generation in 3T3 L1-cells

1995 ◽  
Vol 307 (2) ◽  
pp. 549-556 ◽  
Author(s):  
H I Krieger-Brauer ◽  
H Kather
Keyword(s):  
Growth Factor ◽  
Plasma Membranes ◽  
Platelet Derived Growth Factor ◽  
Fat Cell ◽  
Intact Cells ◽  
Antagonistic Effects ◽  
H2o2 Generation ◽  
Adipose Conversion ◽  
Generating System

3T3 L1-cells, which undergo adipose conversion in vitro, possess a stimulus-sensitive H2O2-generating system in their plasma membrane, and its properties are virtually identical with those of the insulin-sensitive human fat-cell oxidase [Krieger-Brauer and Kather (1992) J. Clin. Invest. 89, 1006-1013]. Insulin and insulin-like growth factor I were found to be active stimulators of NADPH-dependent H2O2 generation. Surprisingly, the acidic (a) and basic (b) isoforms of fibroblast growth factor (FGF) as well as the AA and BB homodimers of platelet-derived growth factor (PDGF) had antagonistic effects on NADPH-dependent H2O2 generation in plasma membranes which were parallelled by corresponding changes in H2O2 accumulation in intact cells. bFGF and PDGF BB (which inhibit NADPH-dependent H2O2 generation) prevented the adipose conversion of 3T3 L1-preadipocytes, and this effect could be reversed by exogenously supplied H2O2. Conversely, aFGF and PDGF AA, which stimulated H2O2 generation, accelerated adipocyte conversion in the presence of insulin and were adipogenic in themselves. Consistently, expression of the adipocyte phenotype induced by insulin, dexamethasone and isobutylmethylxanthine was enhanced in the presence of exogenous hypoxanthine/xanthine oxidase, whereas antioxidants, such as N-acetylcysteine or ascorbate, suppressed the process of differentiation. It is concluded that the H2O2 produced in response to hormones and cytokines may contribute to the development and maintenance of the differentiated state.

scholarly journals A rapid immunological procedure for the isolation of hormonally sensitive rat fat-cell plasma membrane

1976 ◽  
Vol 154 (1) ◽  
pp. 11-21 ◽  
Author(s):  
J P Luzio ◽  
A C Newby ◽  
C N Hales
Keyword(s):  
Plasma Membrane ◽  
Adenylate Cyclase ◽  
Plasma Membranes ◽  
Membrane Preparation ◽  
Fat Cells ◽  
Fat Cell ◽  
Cell Plasma ◽  
Rat Fat Cells ◽  
Plasma Membrane Preparation ◽  
Isolated Rat

1. A rapid method for the isolation of hormonally sensitive rat fat-cell plasma membranes was developed by using immunological techniques. 2. Rabbit anti-(rat erythrocyte) sera were raised and shown to cross-react with isolated rat fat-cells. 3. Isolated rat fat-cells were coated with rabbit anti-(rat erythrocyte) antibodies, homogenized and the homogenate made to react with an immunoadsorbent prepared by covalently coupling donkey anti-(rabbit globulin) antibodies to aminocellulose. Uptake of plasma membrane on to the immunoadsorbent was monitored by assaying the enzymes adenylate cyclase and 5′-nucleotidase and an immunological marker consisting of a 125I-labelled anti-(immunoglobulin G)-anti-cell antibody complex bound to the cells before fractionation. Contamination of the plasma-membrane preparation by other subcellular fractions was also investigated. 4. By using this technique, a method was developed allowing 25-40% recovery of plasma membrane from fat-cell homogenates within 30 min of homogenization. 5. Adenylate cyclase in the isolated plasma-membrane preparation was stimulated by 5 μm-adrenaline.

Regional variation in HDL metabolism in human fat cells: effect of cell size

1987 ◽  
Vol 252 (5) ◽  
pp. E654-E659 ◽  
Author(s):  
J. P. Despres ◽  
B. S. Fong ◽  
P. Julien ◽  
J. Jimenez ◽  
A. Angel
Keyword(s):  
Cell Size ◽  
Cellular Uptake ◽  
Plasma Membranes ◽  
Cholesterol Content ◽  
Cholesterol Concentration ◽  
Fat Cells ◽  
Obese Patients ◽  
Adipocyte Size ◽  
Fat Cell ◽  
Omental Fat

Abdominal obesity is related to reduced plasma high-density lipoprotein (HDL) cholesterol, and both are associated with cardiovascular disease risk. We have observed that plasma membranes from abdominal subcutaneous adipocytes have a greater HDL binding capacity than omental fat cell plasma membranes. The present study examined whether these binding characteristics could be due to differences in fat cell size or cholesterol concentration between the two adipose depots. Abdominal subcutaneous and deep omental fat were obtained from massively obese patients at surgery. Subcutaneous abdominal fat cells were significantly larger and their cellular cholesterol content greater than omental adipocytes. The uptake of HDL by collagenase-isolated fat cells was studied by incubating the cells for 2 h at 37 degrees C with 10 micrograms/ml 125I-HDL2 or 125I-HDL3. In both depots, the cellular uptake of 125I-HDL2 and 125I-HDL3 was specifically inhibited by addition of 25-fold excess unlabeled HDL and a close correlation was observed between the cellular uptake of 125I-HDL2 and 125I-HDL3. In obese patients, the uptake of 125I-HDL was higher in subcutaneous cells than in omental cells [5.85 +/- 0.53 vs. 2.74 +/- 0.30 pmol X 2 h-1. (10(6) cells)-1]. The cellular 125I-HDL uptake was significantly correlated with adipocyte size and fat cell cholesterol content but not with adipocyte cholesterol concentration. These results suggest that the higher HDL uptake observed in subcutaneous cells compared with omental cells in obesity is the result of differences in adipocyte size rather than differences in the cholesterol concentration (cholesterol-to-triglyceride ratio).(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Effect of nutrition on subcellular localization of rat fat-cell lipoprotein lipase

1978 ◽  
Vol 172 (2) ◽  
pp. 239-245 ◽  
Author(s):  
A Vanhove ◽  
C Wolf ◽  
M Breton ◽  
M C Glangeaud
Keyword(s):  
Adipose Tissue ◽  
Lipoprotein Lipase ◽  
Plasma Membranes ◽  
Total Activity ◽  
Normal Diet ◽  
Epididymal Adipose Tissue ◽  
Fat Cells ◽  
Isolated Fat Cells ◽  
Fat Cell ◽  
Intact Tissue

This study supports the possibility for multiple subcellular forms of lipoprotein lipase. 1. The total activity of lipoprotein lipase per g of intact epididymal adipose tissue from fed rats is much higher than that from starved rats. 2. The isolated fat-cells of fed and of starved rats have lipoprotein lipase of almost the same activity per g of fat-pads. The isolated fat-cells of starved rats have a much higher proportion of total activity per g of the intact tissue than do those of fed rats. 3. Under the conditions of homogenization used, only a small proportion of the total activity per g of intact tissue from fed rats was associated with the fat layer which floated to the top of the homogenate during low-speed centrifugation. The different proportions of the specific enzyme activity found in each subcellular fraction are described. 4. Lipoprotein lipase from plasma membranes and microsomal fractions from starved and fed rats was purified by affinity chromatography. 5. The total activity of microsomal lipoprotein lipase per g of intact adipose tissue is enhanced by a normal diet. 6. In intact epididymal adipose tissue from fed rats, the activity per g of tissue of lipoprotein lipase of plasma membranes is much higher than that in the same fraction from starved rats. By contrast, the activities per g of tissue in plasma membranes obtained from starved or from fed rats by collagenase treatment were similar.

scholarly journals Use of a novel rapid preparation of fat-cell plasma membranes employing Percoll to investigate the effects of insulin and adrenaline on membrane protein phosphorylation within intact fat-cells

1980 ◽  
Vol 192 (2) ◽  
pp. 457-467 ◽  
Author(s):  
G J Belsham ◽  
R M Denton ◽  
M J A Tanner
Keyword(s):  
Plasma Membranes ◽  
Microsomal Fraction ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Sodium Dodecyl ◽  
Fat Cells ◽  
Fat Cell ◽  
Rapid Preparation ◽  
Cell Plasma ◽  
Rat Fat Cells

1. A rapid method was developed for the preparation of plasma membranes from either isolated rat fat-cells or intact epididymal fat-pads with the use of density-gradient centrifugation in the presence of Percoll. On the basis of 5′-nucleotidase activity, the yield of plasma membranes was about 50% and purification over 10-fold. Activities of marker enzymes indicated that contamination by mitochondria and microsomal fraction was small. 2. Incorporation of 32Pi into proteins associated with plasma membranes within isolated fat-cells was investigated. Four major bands of labelled phosphoproteins were separated by sodium dodecyl sulphate/polyacrylamide-slab-gel electrophoresis; the apparent subunit mol.wts. were 67 000, 61 000, 26 000 and 20 000. None of these phosphoprotein bands corresponded to periodate/Schiff-staining glycoproteins. The extent of phosphorylation of the 61 000 mol.wt phosphoprotein band was increased by about 30 and 60% after exposure of fat-cells for 15 min to insulin or adrenaline respectively.

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