scholarly journals Affinity purification of cellular retinoic acid-binding protein on 14-carboxy-13-cis-retinamide-sepharose 4B

1989 ◽  
Vol 262 (3) ◽  
pp. 917-922 ◽  
Author(s):  
R K Singh ◽  
B P Sani ◽  
M I Dawson ◽  
Y F Shealy
Keyword(s):  
Retinoic Acid ◽  
Carboxy Group ◽  
Binding Proteins ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Biologically Active ◽  
Amide Bond ◽  
Gel Chromatography ◽  
Fatty Acid Binding ◽  
Sepharose 4B

A biologically active bifunctional retinoid, ethyl 14-carboxyretinoate, has been synthesized and shown to bind cellular retinoic acid (RA)-binding protein (CRABP) via its free carboxy group. We describe herein the synthesis of 14-carboxy-13-cis-retinamide-Sepharose 4B, which is an affinity matrix bearing an all-trans-RA moiety, and thus was used to purify and characterize CRABP from chick-embryo skin. An amide bond was first formed between the free carboxy group of the retinoid and a primary amino group of aminohexyl-Sepharose 4B, by reaction with carbodi-imide, and the ester group of the resin-bound retinoid was then hydrolysed in an alkaline medium. Polyacrylamide-gel electrophoresis and f.p.l.c. Superose column-chromatographic analysis demonstrated that the affinity-purified CRABP (Mr 15,000) was close to electrophoretic homogeneity (greater than 90%) and specifically interacts with RA. By using affinity gel chromatography, conversion of holo-CRABP into apo-CRABP by treatment with p-hydroxymercuribenzoate and a possible involvement of a thiol group in RA binding to CRABP were established. This affinity procedure provides several advantages: (i) 14-carboxy-13-cis-retinamide-Sepharose exhibited high efficiency and selectivity for RA-binding protein (i.e. retinol- or fatty-acid-binding proteins did not bind); (ii) the presence of the amide linkage between the ligand and the matrix makes this affinity resin relatively stable to cytosolic enzymes; and (iii) other RA-binding proteins, e.g. nuclear receptor(s), may be purified.

Properties of triiodothyronine-binding proteins in liver cytosol of rat

1983 ◽  
Vol 61 (9) ◽  
pp. 1035-1041 ◽  
Author(s):  
D. Bellabarba ◽  
S. Bédard ◽  
J. -G. Lehoux
Keyword(s):  
Gel Electrophoresis ◽  
Binding Proteins ◽  
Serum Proteins ◽  
Binding Protein ◽  
Gradient Centrifugation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sedimentation Coefficient ◽  
Gel Chromatography ◽  
Sucrose Density Gradient Centrifugation ◽  
Liver Cytosol

The electrophoretic mobility and the sedimentation coefficient were determined in partially purified preparations of both rat liver cytosol and serum triiodothyronine (T3)-binding proteins. Crude cytosol and serum, each labeled with [125I]T3, were filtered through a Sephadex G-100 column. The cytosol yielded a single T3-binding peak, whereas three binding components were recognized in the serum. Protamine sulfate precipitated the cytosol T3-binding protein, but had no effect on the serum T3-binders. The cytosol protein and the three binding proteins from serum were analyzed by polyacrylamide gel electrophoresis and sucrose density gradient centrifugation. The cytosol binder migrated as a single peak on gel electrophoresis with an Rf of 0.53, whereas the serum proteins had Rfs between 0.27–0.33. The sedimentation coefficient of the cytosol protein was 6.3 S, whereas it was 4.1 S for the major binding protein of the serum. These data indicate that: (i) preliminary purification by gel chromatography is a useful step for better characterization of the T3-binding proteins of the cytosol and serum; (ii) the cytosol binder is an acidic protein with completely different properties from those of the serum T3-binding proteins.

scholarly journals Effects of Simvastatin on Retinoic Acid System in Primary Human Endometrial Stromal Cells and in a Chimeric Model of Human Endometriosis

2013 ◽  
Vol 98 (3) ◽  
pp. E463-E471 ◽  
Author(s):  
Anna Sokalska ◽  
MariaPia Anderson ◽  
Jesus Villanueva ◽  
Israel Ortega ◽  
Kaylon L. Bruner-Tran ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Binding Protein ◽  
Inhibitory Effect ◽  
Experimental Models ◽  
Nude Mouse Model ◽  
Endometrial Stromal Cells ◽  
Fatty Acid Binding ◽  
Proliferation And Apoptosis ◽  
Chimeric Model ◽  
Hes Cells

Context: Retinoic acid (RA) may promote survival or apoptosis of cells, depending on the levels of binding proteins: apoptosis-inducing cellular RA binding protein 2 (CRABP2), and cell survival-promoting fatty acid binding protein 5 (FABP5). Increased cellular uptake of retinol and altered actions of RA related to reduced expression of CRABP2 may contribute to the development of endometriosis. Recently statins have been shown to inhibit growth of human endometrial stromal (HES) cells and to reduce the number and size of endometriotic implants in experimental models of this disorder. Objective: The objective of the study was to determine whether effects of simvastatin on HES cells and experimental endometriotic implants are related to the modulation of the RA system. Methods: Effects of simvastatin and RA on proliferation and apoptosis of HES cells were evaluated. Expression of stimulated by RA 6 (STRA6), CRABP2, and FABP5 was determined by real-time PCR and Western blotting. Effects of simvastatin were also evaluated in a nude mouse model of human endometriosis. Results: Simvastatin potentiated an inhibitory effect of RA on growth of HES cells. In HES cells, simvastatin induced expression of STRA6 and CRABP2 but not FABP5. Similarly, simvastatin treatment of nude mice bearing human endometrial xenografts led to an increased expression of CRABP2 and STRA6 proteins in ectopic lesions. Conclusions: Simvastatin interacts with the RA system, inducing the expression of the key protein regulating the uptake of retinol (STRA6) and the expression of apoptosis-promoting CRABP2. These effects may contribute to cooperative apoptosis-inducing effects of simvastatin and RA and support the examination of these compounds in the treatment of endometriosis.

scholarly journals Is PPARβ/δa Retinoid Receptor?

PPAR Research ◽  
2007 ◽  
Vol 2007 ◽  
pp. 1-5 ◽  
Author(s):  
Daniel C. Berry ◽  
Noa Noy
Keyword(s):  
Retinoic Acid ◽  
Fatty Acid Binding Protein ◽  
Nuclear Translocation ◽  
Retinoic Acid Receptor ◽  
Binding Protein ◽  
Retinoid Receptor ◽  
Expression Ratio ◽  
Fatty Acid Binding ◽  
Bona Fide ◽  
Crabp Ii

The broad ligand-binding characteristic of PPARβ/δhas long hampered identification of physiologically-meaningful ligands for the receptor. The observations that the activity of PPARβ/δis supported by fatty acid binding protein 5 (FABP5), which directly delivers ligands from the cytosol to the receptor, suggest thatbona fidePPARβ/δligands both activate the receptor, and trigger the nuclear translocation of FABP5. Using these criteria, it was recently demonstrated that all-trans-retinoic acid (RA), the activator of the classical retinoic acid receptor RAR, also serves as a ligand for PPARβ/δ. Partitioning of RA between its two receptors was found to be regulated by FABP5, which delivers it to PPARβ/δ, and cellular RA binding protein II (CRABP-II), which targets it to RAR. Consequently, RA activates PPARβ/δin cells that display a high FABP5/CRABP-II expression ratio. It remains to be clarified whether compounds other than RA may also serve as endogenous activators for this highly promiscuous protein.

Adiponectin and adipocyte fatty acid binding protein in the pathogenesis of cardiovascular disease

2012 ◽  
Vol 302 (6) ◽  
pp. H1231-H1240 ◽  
Author(s):  
Aimin Xu ◽  
Paul M. Vanhoutte
Keyword(s):  
Cardiovascular Disease ◽  
Fatty Acid ◽  
Blood Vessels ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Biologically Active ◽  
Adipose Tissues ◽  
Fatty Acid Binding ◽  
Acid Binding Protein ◽  
Obesity And Diabetes

The heart and blood vessels are surrounded by epicardial and perivascular adipose tissues, respectively, which play important roles in maintaining cardiovascular homeostasis by secreting a number of biologically active molecules, termed “adipokines.” Many of these adipokines function as an important component of the ‘adipo-cardiovascular axis’ mediating the cross talk between adipose tissues, the heart, and the vasculature. On the one hand, most adipokines [including tumor necrosis factor-α, resistin, adipocyte fatty acid binding protein (A-FABP), and lipocalin-2] are proinflammatory and causally associated with endothelial and cardiac dysfunction by their endocrine/paracrine actions. On the other hand, adiponectin is one of the few adipokines that possesses multiple salutary effects on the prevention of cardiovascular disease, because of its pleiotropic actions on the heart and the blood vessels. The discordant production of adipokines in dysfunctional adipose tissue is a key contributor to obesity-related cardiovascular disease. This review provides an update in understanding the roles of adipokines in the pathogenesis of cardiovascular disorders associated with obesity and diabetes and focuses on the two most abundant adipokines, adiponectin and A-FABP. Indeed, data from both animal studies and clinical investigations imply that these two adipokines are prognostic biomarkers for cardiovascular disease and even promising therapeutic targets for its treatment.

scholarly journals Comparison of the binding affinities of acyl-CoA-binding protein and fatty-acid-binding protein for long-chain acyl-CoA esters

1990 ◽  
Vol 265 (3) ◽  
pp. 849-855 ◽  
Author(s):  
J T Rasmussen ◽  
T Börchers ◽  
J Knudsen
Keyword(s):  
Fatty Acid ◽  
Binding Proteins ◽  
Fatty Acid Binding Protein ◽  
Binding Assay ◽  
Binding Protein ◽  
Fatty Acid Binding Proteins ◽  
Binding Affinities ◽  
Fatty Acid Binding ◽  
Chain Acyl ◽  
Long Chain

Bovine and rat liver acyl-CoA-binding proteins (ACBP) were found to exhibit a much higher affinity for long-chain acyl-CoA esters than both bovine hepatic and cardiac fatty-acid-binding proteins (hFABP and cFABP respectively). In the Lipidex 1000- as well as the liposome-binding assay, bovine and rat hepatic ACBP effectively bound long-chain acyl-CoA ester, h- and c-FABP were, under identical conditions, unable to bind significant amounts of long-chain acyl-CoA esters. When FABP, ACBP and [1-14C]hexadecanoyl-CoA were mixed, hexadecanoyl-CoA could be shown to be bound to ACBP only. The experimental results give strong evidence that ACBP, and not FABP, is the predominant carrier of acyl-CoA in liver.

scholarly journals Studies on fatty acid-binding proteins. Changes in the concentration of hepatic fatty acid-binding protein during development in the rat

1987 ◽  
Vol 242 (3) ◽  
pp. 919-922 ◽  
Author(s):  
M Sheridan ◽  
T C I Wilkinson ◽  
D C Wilton
Keyword(s):  
Fatty Acid ◽  
Binding Proteins ◽  
Fatty Acid Binding Protein ◽  
Fluorescence Enhancement ◽  
Binding Protein ◽  
Fold Increase ◽  
Fatty Acid Binding Proteins ◽  
Fatty Acid Binding ◽  
Acid Binding Protein ◽  
Hepatic Fatty Acid

The concentration of hepatic fatty acid-binding protein was determined in the livers of rats at various stages of development from foetus to young adult. Fatty acid-binding protein concentrations were determined by quantifying the fluorescence enhancement on the binding of the fluorescent probe 11-(dansylamino)-undecanoic acid. A 20-fold increase in the concentration of the protein was observed between the foetus and adult, and this increase was confirmed by immuno-blotting. No other protein in the 14,000-Mr range was observed in the foetus. Possible alternative fatty acid-binding proteins could not be detected in h.p.l.c.-fractionated foetal cytosol by the fluorescence-enhancement method.

scholarly journals Selective Cooperation between Fatty Acid Binding Proteins and Peroxisome Proliferator-Activated Receptors in Regulating Transcription

2002 ◽  
Vol 22 (14) ◽  
pp. 5114-5127 ◽  
Author(s):  
Nguan-Soon Tan ◽  
Natacha S. Shaw ◽  
Nicolas Vinckenbosch ◽  
Peng Liu ◽  
Rubina Yasmin ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Fatty Acid ◽  
Binding Proteins ◽  
Lipid Binding ◽  
Fatty Acid Binding Proteins ◽  
Peroxisome Proliferator ◽  
Fatty Acid Binding ◽  
Peroxisome Proliferator Activated Receptors ◽  
Retinoic Acid Binding Proteins ◽  
Crabp Ii

ABSTRACT Lipophilic compounds such as retinoic acid and long-chain fatty acids regulate gene transcription by activating nuclear receptors such as retinoic acid receptors (RARs) and peroxisome proliferator-activated receptors (PPARs). These compounds also bind in cells to members of the family of intracellular lipid binding proteins, which includes cellular retinoic acid-binding proteins (CRABPs) and fatty acid binding proteins (FABPs). We previously reported that CRABP-II enhances the transcriptional activity of RAR by directly targeting retinoic acid to the receptor. Here, potential functional cooperation between FABPs and PPARs in regulating the transcriptional activities of their common ligands was investigated. We show that adipocyte FABP and keratinocyte FABP (A-FABP and K-FABP, respectively) selectively enhance the activities of PPARγ and PPARβ, respectively, and that these FABPs massively relocate to the nucleus in response to selective ligands for the PPAR isotype which they activate. We show further that A-FABP and K-FABP interact directly with PPARγ and PPARβ and that they do so in a receptor- and ligand-selective manner. Finally, the data demonstrate that the presence of high levels of K-FABP in keratinocytes is essential for PPARβ-mediated induction of differentiation of these cells. Taken together, the data establish that A-FABP and K-FABP govern the transcriptional activities of their ligands by targeting them to cognate PPARs in the nucleus, thereby enabling PPARs to exert their biological functions.

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