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1.
The simultaneous determination of heavy metals in microsamples of chromatographically isolated cadmium-binding protein (Cd-BP) from rat liver was performed by neutron activation analysis. The results suggested that metals other than those already reported (Cd, Zn, Cu, and Hg) can bind the protein. These observations were confirmed by in vivo radiotracer experiments by injecting i.p. 21 labelled metal ions in cadmium-treated rats. Of the metals tested, 109Cd, 65Zn, 64Cu, 203Hg, 106Ag and 113Sn were found incorporated in the Cd-BP. The incorporation of 35S-cysteine, used as an indicator of Cd-BP biosynthesis, was increased in rats exposed to cadmium as compared to untreated animals. In order to establish the influence of other metal ions on the biosynthesis of Cd-BP and the incorporation of cadmium in the protein, in vivo experiments were carried out by i.p. injection of 109Cd and 35S-cysteine. In the presence of 42 metal ions no influence was observed on the incorporation of the two radioisotopes in the Cd-BP. These observations tend to support the hypothesis that cadmium can act as a highly specific inducer of Cd-BP and that this protein might be involved in the metabolism of several heavy metals. 相似文献
2.
G L Catignani 《Biochemical and biophysical research communications》1975,67(1):66-72
Gel filtration and sucrose density gradient analysis of rat liver high speed supernatant revealed the presence of a protein capable of binding [3H] α-tocopherol. The protein sedimented with an S value of 3.0. Gel filtration yielded an estimated molecular weight of 31,000. Specificity for α-tocopherol was demonstrated by competition for binding of [3H]α-tocopherol with unlabeled α-tocopherol, but not with α-tocopheryl quinone or α-tocopheryl acetate. Pronase digestion completely abolished binding. 相似文献
3.
Estrogen binding sitesin the embryonic chicken liver 总被引:1,自引:0,他引:1
M Gschwendt 《FEBS letters》1977,75(1):272-276
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Testosterone binding protein from rat liver cytosol, which had been incubated with [3H]testosterone followed by treatment with dextran-coated charcoal, was analyzed by DEAE-cellulose and phosphocellulose chromatography. On DEAE-cellulose chromatography, two distinct peaks of radioactivity were eluted at 0.07 M and 0.19 M KCl, both sedimented in 4 S regions. Phosphocellulose chromatography resulted in a broad peak at 0.08 M KCl, with a shoulder at 0.04 M KCl, both sedimented at 4 S. These findings indicated that testosterone binding protein consists of two types of components each with 4 S. 相似文献
5.
A binding protein with apparent specificity for beta-glucuronidase has been partially purified from a Triton X-100 extract of rat liver microsomes by affinity chromatography on glucuronidase-Sepharose 2B. It appears that once removed from the membrane, this binding protein self-aggregates to form large macromolecular complexes. With the use of polyacrylamide gel electrophoretic and sucrose density gradient ultracentrifugation assays to monitor the conversion of glucuronidase tetramer to a very high molecular weight complex, it was shown that the binding activity is heatlabile and protease-sensitive. However, binding activity is not influenced by salts, carbohydrates, other proteins or glycoproteins, or by extensive periodate oxidation of beta-glucuronidase, nor does binding occur with any other protein tested. The binding protein does not discriminate against any form of beta-glucuronidase from any rat organ tested. However, the binding protein does show organ localization, being present in the liver and kidney but not the spleen. The possible relationship of this binding protein to egasyn, a membrane protein which stabilizes beta-glucuronidase in mouse liver endoplasmic reticulum, is discussed. 相似文献
6.
Control of cadmium binding protein synthesis in rat liver 总被引:8,自引:0,他引:8
7.
A retinylphosphate binding activity, resolved during purification, has been discovered in rat liver cytosol. The partial purification includes ammonium sulfate precipitation and DEAE-cellulose chromatography. The macromolecular component responsible for the binding has a sedimentation coefficient of about 2 S and is sensitive to pronase. This binding is reversible and specific for retinylphosphate, since retinol, retinoic acid and retinoylphosphate do not compete with [3H]retinylphosphate. 相似文献
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Estrogen receptor in rat liver and its dependence on prolactin. 总被引:8,自引:0,他引:8
Estrogen receptor is shown to be present in the livers of adult rats. The receptor binds estradiol-17beta with a Kd of 1 x 10(-10) M and sediments at 8 S in sucrose gradients. Other estrogens and anti-estrogens compete for estradiol binding, while nonestrogenic steroids do not. Receptor levels fall dramatically after hypophysectomy, but can be partially restored within 18 hours by a single injection of prolactin. It is known that prolactin critically regulates the level of its own receptor in the liver, and we now suggest that it also exerts a primary control over the availability of liver estrogen receptor. 相似文献
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In this study, lysophosphatidylcholine (lysoPC) was shown to bind to a fatty acid binding protein isolated from rat liver. To demonstrate the binding, lysoPC was incorporated into multilamellar liposomes and incubated with protein. For comparison, binding of both lysoPC and fatty acid to liver fatty acid binding protein, albumin, and heart fatty acid binding protein were measured. At conditions where palmitic acid bound to liver fatty acid binding protein and albumin at ligand to protein molar ratios of 2:1 and 5:1, respectively, lysoPC binding occurred at molar ratios of 0.4:1 and 1:1. LysoPC did not bind to heart fatty acid binding protein under conditions where fatty acid bound at a molar ratio of 2:1. Competition experiments between lysoPC and fatty acid to liver fatty acid binding protein indicated separate binding sites for each ligand. An equilibrium dialysis cell was used to demonstrate that liver fatty acid binding protein was capable of transporting lysoPC from liposomes to rat liver microsomes, thereby facilitating its metabolism. These studies suggest that liver fatty acid binding protein may be involved in the intracellular metabolism of lysoPC as well as fatty acids, and that functional differences may exist between rat liver and heart fatty acid binding protein. 相似文献
12.
A low-molecular-weight protein from rat liver that resembles ligandin in its binding properties. 总被引:4,自引:6,他引:4
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A protein of S20,W 1.6S and mol.wt. 14000, which binds covalently a metabolite of the aminoazodye carcinogen NN-dimethyl-4-amino-3'-methylazobenzene, was isolated from rat liver cytosol from both carcinogen-treated and normal rats. The protein binds non-covalently palmitoyl-CoA, fatty acids, bilirubin, sex steroids and their sulphates, bile acids and salts, bromosulphophthalein, diethylstilboestrol and 20-methylcholanthrene with a wide range of affinities. The protein is isolated as three components with isoelectric points of 5.0, 5.9 and 7.6 by a method involving isoelectric focusing. All three components have closely similar amino acid analyses, tryptic-peptide 'maps' and u.v. spectra. Each single component redistributes into all three on further electrophoresis. However, the three forms differ in their binding characteristics, the form of pI 7.6 having much the highest affinity for compounds bound non-covalently. The protein was identified immunologically in rat liver, small intestine, adipose tissue, skeletal muscle, myocardium and testis. The protein was compared with other hepatic binding-protein preparations of similar molecular weight. 相似文献
13.
Effect of liver fatty acid binding protein on fatty acid movement between liposomes and rat liver microsomes. 总被引:3,自引:0,他引:3
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Although movement of fatty acids between bilayers can occur spontaneously, it has been postulated that intracellular movement is facilitated by a class of proteins named fatty acid binding proteins (FABP). In this study we have incorporated long chain fatty acids into multilamellar liposomes made of phosphatidylcholine, incubated them with rat liver microsomes containing an active acyl-CoA synthetase, and measured formation of acyl-CoA in the absence or presence of FABP purified from rat liver. FABP increased about 2-fold the accumulation of acyl-CoA when liposomes were the fatty acid donor. Using fatty acid incorporated into liposomes made either of egg yolk lecithin or of dipalmitoylphosphatidylcholine, it was found that the temperature dependence of acyl-CoA accumulation in the presence of FABP correlated with both the physical state of phospholipid molecules in the liposomes and the binding of fatty acid to FABP, suggesting that fatty acid must first desorb from the liposomes before FABP can have an effect. An FABP-fatty acid complex incubated with microsomes, in the absence of liposomes, resulted in greater acyl-CoA formation than when liposomes were present, suggesting that desorption of fatty acid from the membrane is rate-limiting in the accumulation of acyl-CoA by this system. Finally, an equilibrium dialysis cell separating liposomes from microsomes on opposite sides of a Nuclepore filter was used to show that liver FABP was required for the movement and activation of fatty acid between the compartments. These studies show that liver FABP interacts with fatty acid that desorbs from phospholipid bilayers, and promotes movement to a membrane-bound enzyme, suggesting that FABP may act intracellularly by increasing net desorption of fatty acid from cell membranes. 相似文献
14.
Studies on fatty acid-binding proteins. The binding properties of rat liver fatty acid-binding protein.
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The inactive 2Fe species of the Fe protein of the nitrogenase of Klebsiella pneumoniae was generated by treating oxidized Fe protein (Kp2) with MgATP and chelator. Incubation of the 2Fe species of Kp2 with the sulphurtransferase rhodanese in the presence of thiosulphate, ferric citrate and reduced lipoate reproducibly restored activity. The extent of restoration of activity depended on the molar ratio of 2Fe Kp2 to rhodanese and was time-dependent. Re-activation did not occur in the reaction mixture lacking rhodanese. 相似文献
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Hyaluronic acid binding protein (HBP) was purified homogeneously from normal adult rat liver by hyaluronate-sepharose affinity chromatography. The molecular weight of this protein as determined by gel filtration was found to be 64,000 daltons. This protein HBP appeared as a single band in non-dissociating gel electrophoresis and has a subunit of molecular weight approximately 12,000 as determined by SDS-gel electrophoresis. 相似文献
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Zimmermann EM Li L Hoyt EC Pucilowska JB Lichtman S Lund PK 《American journal of physiology. Gastrointestinal and liver physiology》2000,278(3):G447-G457
The liver is a major source of circulating insulin-like growth factor I (IGF-I), and it also synthesizes several classes of IGF binding proteins (IGFBPs). Synthesis of IGF-I and IGFBPs is regulated by hormones, growth factors, and cytokines. They are nutritionally regulated and expressed in developmentally specific patterns. To gain insight into cellular regulatory mechanisms that determine hepatic synthesis of IGF-I and IGFBPs and to identify potential target cells for IGF-I within the liver, we studied the cellular sites of synthesis of IGF-I, IGF receptor, growth hormone (GH) receptor, and IGFBPs in freshly isolated rat hepatocytes, endothelial cells, and Kupffer cells. We also localized cellular sites of IGFBP synthesis by in situ hybridization histochemistry. Western ligand and immunoblot analyses were used to determine IGFBP secretion by isolated cells. Two IGF-I mRNA subtypes with different 5' ends (class 1 and class 2) were detected in all isolated liver cell preparations. Type 1 IGF receptor mRNA was detected in endothelial cells, indicating that these cells are a local target for IGF actions in liver. GH receptor was expressed in all cell preparations, consistent with GH regulation of IGF-I and IGFBP synthesis in multiple liver cell types. The IGFBPs expressed striking cell-specific expression. IGFBP-1 was synthesized only in hepatocytes, and IGFBP-3 was expressed in Kupffer and endothelial cells. IGFBP-4 was expressed at high levels in hepatocytes and at low levels in Kupffer and endothelial cells. Cell-specific expression of distinct IGFBPs in the liver provides the potential for cell-specific regulation of hepatic and endocrine actions of IGF-I. 相似文献
19.
A radioimmunoassay has been developed for the folate binding protein from rat liver cytosol with a molecular weight of 150,000 which was recently purified to homogeneity (Suzuki, N., and Wagner, C., 1980, Arch. Biochem. Biophys.199, 236–248). This method has indicated that the binding protein (FBP-CII) is found primarily in the liver. A significant amount of FBP-CII was also found in the kidney and much reduced levels in spleen, serum, brain, lung, and heart. No FBP-CII could be detected in small intestine, skeletal muscle, or testes. Small amounts of cross-reacting material were found in the livers of mouse, dog, chick, and humans. Levels of FBP-CII were not decreased in the livers of folate-deficient rats. Assays of rat fetal liver and kidney 2 days prior to birth showed much lower levels which increased rapidly at birth. These data are consistent with the FBP-CII fulfilling a role as a folate storage protein in rat liver. 相似文献
20.
Studies were performed to elucidate the ontogeny of a single class of androgen binding protein in male rat liver cytosol which exhibits characteristics of a ligand specific, high affinity (Kd = 2.3 nM), 8S-receptor capable of nuclear translocation. Detectable levels of receptor first appear at 45 days of age in the male and reach maximum concentration at 65 days. Barely detectable levels are seen in females throughout the duration of study (80 days). Gonadectomy in both sexes (65 days) and androgen treatment of oophorectomized females do not alter the normal development of sexual differentiation of the high affinity androgen receptor. After neonatal castration (2 days) and DES replacement however, receptor sites do not undergo differentiation and adult males exhibit female levels. Conversely, neonatal androgen replacement in 2-day castrates partially restores the level of binding sites to control males values (TP, 71%; DHT, 51%). Neonatal castration without replacement retards but does not fully eliminate sexual differentiation of levels of receptor sites in adult males. Likewise, neonatal androgen treatment in females results in a partial masculinization of binding sites. Following hypophysectomy, levels of receptor sites in females are similar to intact or hypophysectomized males; sexual differences in the adult are abolished. These studies suggest that sexual differentiation of specific liver cytosol androgen binding sites in the adult may be partially programmed at birth by testicular androgen and furthermore, adult sexual dimorphism is maintained through an inhibitory influence of the pituitary in the female. 相似文献