Soy protein diets lower plasma cholesterol in hyperlipoproteinemic human subjects, as well as in animal models. We fed 7-wk-old male obese (fa/fa) and lean Zucker rats a modified AIN-76 diet (20 g protein/kg diet) containing casein (C), low isoflavone soy protein (38 mg isoflavones/kg diet; LI), or high isoflavone soy protein (578 mg isoflavones/kg diet; HI) for 70 d. In obese rats, plasma total cholesterol was 21 and 29% lower in the LI and HI groups, respectively, than in the C group (P: = 0.004). Liver weight and liver triglyceride and cholesteryl ester concentrations were 27, 33 and 46% lower, respectively, in the LI group than in the C group (P: < 0.003). These liver measurements were 23, 24 and 57% lower, respectively, in the HI group than in the LI group (P: < 0.05). In a complementary study, 5-wk-old male Sprague-Dawley rats were fed the same C, LI and HI diets for 42 d. Thrombin-mediated platelet serotonin release in vitro was 13% lower in the HI group than in the C group (P: = 0.003). In a third study, 7-wk-old male Sprague-Dawley rats were fed either a modified AIN-76 control diet or a high fat casein-based atherogenic diet (140 g fat, 12 g cholesterol, and 2 g cholic acid/kg diet) with or without a soy isoflavones extract (983 mg isoflavones/kg diet) for 63 d. Addition of the isoflavones extract to the atherogenic diet lowered the liver triglyceride concentration by 33% relative to the atherogenic diet without isoflavones (P: = 0.0001). Our studies suggest that the hypocholesterolemic mechanism of dietary soy protein involves a cooperative interaction between the protein and isoflavone-enriched fraction that lowers hepatic lipid concentrations. We speculate that modulation of liver and plasma lipid homeostasis can also lower blood platelet sensitivity.
Chemical and proteolytic digestion of intact erythrocyte glucose transporter as well as purified transporter protein has been used to localize the derivatization site for the photoaffinity agent 3-[125I]iodo-4-azido-phenethylamino-7-O-succinyldeacetylforskol in [( 125I]IAPS-forskolin). Comparison of the partial amino acid sequence of the labelled 18 kDa tryptic fragment with the known amino acid sequence for the HepG2 glucose transporter confirmed that the binding site for IAPS-forskolin is between the amino acid residues Glu254 and Tyr456. Digestion of intact glucose transporter with Pronase suggests that this site is within the membrane bilayer. Digestion of labelled transporter with CNBr generated a major radiolabelled fragment of Mr approximately 5800 putatively identified as residues 365-420. Isoelectric focusing of Staphylococcus aureus V8 proteinase-treated purified labelled tryptic fragment identified two peptides which likely correspond to amino acid residues 360-380 and 381-393. The common region for these radiolabelled peptides is the tenth putative transmembrane helix of the erythrocyte glucose transporter, comprising amino acid residues 369-389. Additional support for this conclusion comes from studies in which [125I]APS-forskolin was photoincorporated into the L-arabinose/H(+)-transport protein of Escherichia coli. Labelling of this transport protein was protected by both cytochalasin B and D-glucose. The region of the erythrocyte glucose transporter thought to be derivatized with IAPS-forskolin contains a tryptophan residue (Trp388) that is conserved in the sequence of the E. coli arabinose-transport protein.
We examined the effects of diets based on a low isoflavone or a high isoflavone soy protein isolates in normal, growth-hormone receptor knockout and Ames dwarf, and Prop 1 (df) mice that are hypoinsulinemic, insulin-sensitive, and exceptionally long-lived, as well as in growth hormone transgenic mice that are hyperinsulinemic, insulin-resistant, dyslipidemic, and short-lived. Soybean diets tended to normalize plasma cholesterol levels in dwarf and transgenic mice, while low isoflavone diet reduced plasma triglycerides in most of the examined genotypes. The effects of low isoflavone and high isoflavone diets on the levels of free and esterified cholesterol in the liver were strongly genotype-dependent. Fasting blood glucose levels were reduced and glucose tolerance improved by both low isoflavone and high isoflavone diets in growth hormone-transgenic mice and in their normal siblings. Glucose tolerance was also improved by high-isoflavone diet in growth hormone receptor knockout mice. Lifespan was increased by low isoflavone diet in normal mice from two of the examined stocks. High isoflavone diet increased lifespan in normal animals from one line, but reduced lifespan of normal mice from a different line. We conclude that dietary soy protein intake can improve plasma and hepatic lipid profiles, reduce fasting glucose, enhance capacity for glucose tolerance, and prolong life, but all of these effects are strongly genotype-dependent.
The tryptophan residues 388 and 412 in the glucose transporter GLUT1 were altered to leucine (L) by site-directed mutagenesis and were transiently expressed in COS-7 cells. As assessed by immunoblotting, comparable numbers of glucose transporters were present in plasma membranes from cells transfected with wild-type GLUT1, GLUT1-L388 or GLUT1-L412. Transfection of the wild-type GLUT1 gave rise to a 3-fold increase in the reconstituted glucose transport activity recovered from plasma membranes. In contrast, transfection of GLUT1-L412 failed to increase the reconstituted transport activity, whereas transfection of GLUT1-L388 produced only a 70% increase. Photolabelling of GLUT1-L412 with 3-[125I]iodo-4-azidophenethylamido-7-O-succinyldeacetyl (125IAPS)-forskolin was not different from that of the wild-type GLUT1, whereas the GLUT1-L388 incorporated 70% less photolabel than did the wild-type GLUT1. These data suggest a dissociation of the binding sites of forskolin and glucose in GLUT1. Whereas both tryptophan-388 and tryptophan-412 appear indispensable for the function of the transporter, only tryptophan-388 is involved in the binding of the inhibitory ligand forskolin.
The photoincorporation of cytochalasin B into the human erythrocyte glucose transporter and purified G-actin previously reported by this laboratory [Shanahan, M.F. (1982) J. Biol. Chem. 257, 7290-7293] was investigated. [3H]Cytochalasin B photolabeled polypeptides of Mr approximately 43,000-73,000, as determined by polyacrylamide gel electrophoresis, in a concentration-dependent manner with maximum incorporation occurring at 5 microM [3H]cytochalasin B and a half-maximum value of 0.63 microM. This incorporation, previously shown to be partially blocked in the presence of D- but not L-glucose, did not occur in the absence of photolysis and increased linearly with a photolysis time up to 30 s. The reaction was relatively insensitive to pH in the range of pH 6-9, but apparent non-specific labeling significantly increased at pH 5. The effect of cytochalasin B photoincorporation on D-glucose uptake in intact erythrocytes was also examined. Purified chicken muscle F-actin was also photolabeled with this ligand, but at a specific activity of incorporation (pmol/mg of protein) approximately 50 times lower than that of the erythrocyte transporter polypeptides. D-Glucose had no effect on this incorporation while 10(-4) M cytochalasin E completely blocked actin photolabeling. The efficiency of photoincorporation for both the transporter and F-actin was around 1%. Extraction of [3H]cytochalasin B labeled membranes with Triton X-100 resulted in the selective elution of labeled polypeptides from the transporter region while cytochalasin B labeled polypeptides in the region of red cell actin remained in the extracted pellet.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.