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1.
Michaël M. Vork Jan F. C. Glatz Ger J. van der Vusse 《Molecular and cellular biochemistry》1993,123(1-2):175-184
To obtain insight into the relation between the release of heart-type fatty acid-binding protein (H-FABPc) and of long-chain fatty acids (FA) from injured cardiac tissue, rat hearts were Langendorff perfused according to the following scheme: 30 min normoxia, 60 min ischemia, 30 min reperfusion, 10 min Ca2+ free perfusion and finally 10 min Ca2+ repletion. During this protocol right ventricular (Q
rv
) and interstitial effluent samples (Q
i
) were collected at regular intervals. During reperfusion a total of 0.8±0.1 nmol H-FABPc but no FA were detected in the effluents. However, during Ca2+ readmission, 45±4 nmol H-FABPc (80–90% of total tissue content) was released with an initial (first 3 min) simultaneous release of FA (FA/H-FABPc ratio 0.90±0.07 mol/mol). Thereafter, FA release continued at 10–15 nmol per min mainly inQ
rv
while the rate of H-FABPc release decreased. During Ca2+ repletion, tissue FA content raised rapidly from 168±20 to 1918±107 nmol/g dry weight. These findings suggest that after severe cardiac damage initially FA is released bound to H-FABPc, whereas further FA release occurs in a non-protein bound manner. 相似文献
2.
Summary At least three different proteins are implicated in the cellular transport of fatty acid moieties: a plasmalemmal membrane and a cytoplasmic fatty acid-binding protein (FABPPM and FABPC, respectively) and cytoplasmic acyl-CoA binding protein (ACBP). Their putative main physiological significance is the assurance that long-chain fatty acids and derivatives, either in transit through membranes or present in intracellular compartments, are largely complexed to proteins. FABPC distinguishes from the other proteins in that distinct types of FABPC are found in remarkable abundance in the cytoplasmic compartment of a variety of tissues. Although their mechanism of action is not yet fully elucidated, current knowledge suggests that the function of this set of proteins reaches beyond simply aiding cytoplasmic solubilization of hydrophobic ligands, but that they can be assigned several regulatory roles in cellular lipid homeostasis. 相似文献
3.
Judith Storch 《Molecular and cellular biochemistry》1990,98(1-2):141-147
Summary Fatty acid-binding proteins (FABP) are distinct but related gene products which are found in many mammalian cell types. They are generally present in high abundance, and are found in those tissues where free fatty acid (ffa) flux is high. The function(s) of FABP is unknown. Also not known is whether all FABP function similarly in their respective cell types, or whether different FABP have unique functions. The purpose of these studies was to assess whether different members of the FABP family exhibit different structural and functional properties. Two fluorescent analogues of ffa were used to compare the liver (L-FABP) and heart (H-FABP) binding proteins. The propionic acid derivative of diphenylhexatriene (PADPH) was used to examine the physical properties of the ffa binding site on L- and H-FABP, as well as the relative distribution of ffa between FABP and membranes. An anthroyloxy-derivative of palmitic acid, 2AP, was used to monitor the transfer kinetics of ffa from liver or heart FABP to acceptor membranes, using a resonance energy transfer assay. The results demonstrate that the ffa binding sites of both FABP are hydrophobic in nature, although the L-FABP site is more nonpolar than the H-FABP site. Equilibration of PADPH between L-FABP and phosphatidylcholine (PC) bilayers resulted in a molar partition preference of > 20: 1, L-FABP : PC. Similar studies with H-FABP resulted in a PADPH partition preference of only 3:1, H-FABP : PC. Finally, the transfer of 2AP from H-FABP to acceptor membranes was found to be 50-fold faster than transfer from L-FABP. These studies demonstrate that important structural and functional differences exist between different members of the FABP family, and therefore imply that the roles of different FABP may be unique.Abbreviations FABP
Fatty Acid-Binding Protein
- L-FABP
Liver FABP
- H-FABP
Heart FABP
- SUV
Small Unilamellar Vesicle
- PADPH
3-[p-(6-Phenyl)-1,3,5-Hexatrienyl]-phenylpropionic acid
- 2AP
2-(9-Anthroyloxy)Palmitic acid
- Q
Quantum yield
- F
Fluorescence lifetime 相似文献
4.
The mammalian fatty acid-binding proteins (FABPs) are thought to be important for the transport and metabolism of fatty acids in numerous cell types. The transfer of FA from different members of the FABP family to membranes has been shown to occur by two distinct mechanisms, an aqueous diffusion-based mechanism and a collisional mechanism, wherein the FABP interacts directly with membrane acceptors. Much of the work that underlies this concept comes from efforts using rodent FABPs. Given the increasing awareness of links between FABPs and several chronic diseases in humans, it was important to establish the mechanisms of FA transfer for human FABPs. In the present studies, we examined the rate and mechanism of fatty acid transfer from four pairs of human and rodent (rat or mouse, as specified) FABPs: hLFABP and rLFABP, hIFABP and rIFABP, hHFABP and rHFABP, and hAFABP and mAFABP. In the case of human IFABP, both the Ala54 and Thr54 forms were examined. The results show clearly that for all FABPs examined, the mechanisms of ligand transfer observed for rodent proteins hold true for their human counterparts. Moreover, it appears that the Ala to Thr substitution at residue 54 of the human IFABP does not alter the fundamental mechanism of ligand transfer to membranes, but nevertheless causes a consistent decrease in the rate of transfer. 相似文献
5.
Gerhard E. Gerber Dev Mangroo Bernardo L. Trigatti 《Molecular and cellular biochemistry》1993,123(1-2):39-44
A photoaffinity labeling method was developed to identify and characterize high affinity fatty acid-binding proteins in membranes. The specific labeling of these sites requires the use of low concentrations (nanomolar) of the photoreactive fatty acid 11-m-diazirinophenoxy-[11-3H]undecanoate. It was delivered as a bovine serum albumin (BSA) complex which serves as a reservoir for fatty acid and thus allows precise control of unbound fatty acid concentrations. ThefadL protein ofE. coli, which is required for fatty acid permeation of its outer membrane, was labeled by the photoreactive fatty acid neither specifically nor saturably when the probe was added in the absence of BSA; however when a nanomolar concentration of the uncomplexed probe was maintained in the presence of BSA, the labeling of thefadL protein was highly specific and saturable. This photoaffinity labeling method was also used to characterize a 22 kDa, high affinity fatty acid-binding protein which we have recently identified in the plasma membrane of 3T3-L1 adipocytes. This protein bound the probe with a Kd of 216 nM. The approach described is easily capable of identifying membrane-bound fatty acid-binding proteins and can distinguish between those of high and low affinities for fatty acids. It represents a general method for the identification and characterization of fatty acid-binding proteins.Abbreviations BSA
Bovine Serum Albumin
- DAP
m-Diazirinophenoxy
- SDS-PAGE
Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis 相似文献
6.
Dick J. van der Horst Jan M. van Doorn Paul C. C. M. Passier Michael M. Vork Jan F. C. Glatz 《Molecular and cellular biochemistry》1993,123(1-2):145-152
Since insect flight muscles are among the most active muscles in nature, their extremely high rates of fuel supply and oxidation pose interesting physiological problems. Long-distance flights of species like locusts and hawkmoths are fueled through fatty acid oxidation. The lipid substrate is transported as diacylglycerol in the blood, employing a unique and efficient lipoprotein shuttle system. Following diacylglycerol hydrolysis by a flight muscle lipoprotein lipase, the liberated fatty acids are ultimately oxidized in the mitochondria. Locust flight muscle cytoplasm contains an abundant fatty acid-binding protein (FABP). The flight muscle FABP ofLocusta migratoria is a 15 kDa protein with an isoelectric point of 5.8, binding fatty acids in a 1:1 molar stoichiometric ratio. Binding affinity of the FABP for longchain fatty acids (apparent dissociation constant Kd=5.21±0.16 M) is however markedly lower than that of mammalian FABPs. The NH2-terminal amino acid sequence shares structural homologies with two insect FABPs recently purified from hawkmoth midgut, as well as with mammalian FABPs. In contrast to all other isolated FABPs, the NH2 terminus of locust flight muscle FABP appeared not to be acetylated. During development of the insect, a marked increase in fatty acid binding capacity of flight muscle homogenate was measured, along with similar increases in both fatty acid oxidation capacity and citrate synthase activity. Although considerable circumstantial evidence would support a function of locust flight muscle FABP in intracellular uptake and transport of fatty acids, the finding of another extremely well-flying migratory insect, the hawkmothAcherontia atropos, which employs the same lipoprotein shuttle system, however contains relatively very low amounts of FABP in its flight muscles, renders the proposed function of FABP in insect flight muscles questionable. 相似文献
7.
Summary A variety of designations is currently being used to refer to cellular fatty acid-binding proteins (FABPs). Besides from the use of other general names (e.g. Z protein), confusion mostly arises from the application of various abbreviations and symbols to denote the tissue(s) of origin and cellular localization (cytoplasm, plasma membrane) of a specific FABP. In order to minimize confusion a more unified and rational nomenclature is proposed, which is based on application of the formula X-FABPy. The prefix X is a capital letter indicating the tissue of greatest abundance, the suffix Y similarly denotes the (sub)cellular localization of the protein. The general and functional name fatty acid-binding protein (FABP) is preferred for the cellular proteins with the property to bind fatty acids, unless future research reveals that the binding of fatty acids is not the primary biological property or physiological role of (some of) these proteins. 相似文献
8.
Randall M. Jones M. Renuka Prasad Dipak K. Das 《Molecular and cellular biochemistry》1990,98(1-2):161-166
Summary In this study, we examined the effects of exposure of heart fatty acid-binding protein (h-FABP) to chemically generated O2
– or OH · with respect to its oleate binding and to its electrophoretic properties. Purified rat h-FABP at 40 M scavenged as much as 30% O2
– and 85% OH ·. On the other hand, when 2 nmol (4 M) FABP were exposed to free radicals, the maximum oleate binding capacity as measured by Scatchard analysis was reduced only by 14% and 27% for O2
– and OH ·, respectively. The electrophoretic pattern of free radical-exposed FABP was not markedly different when examined either by the non-denaturing or by denaturing PAGE, suggesting the absence of any degradation or aggregation of FABP by O2
– or OH ·. It is hypothesized that O2
– or OH · in physiological concentration may not alter the function of FABP markedly in the ischemic-reperfused myocardium.Abbreviations h-FABP
Heart Fatty Acid Binding Protein
- NEFA
Non-Esterified Fatty Acids
- O2
–
Superoxide anions
- OH·
hydroxyl radicals
- OCI
hypohalite radicals
- H2O2
hydrogen peroxide
- HPLC
High Pressure Liquid Chromatography 相似文献
9.
Jan F. C. Glatz Michaël M. Vork Ger J. van der Vusse 《Molecular and cellular biochemistry》1993,123(1-2):167-173
Ischemia of the heart is accompanied by the tissue accumulation of long-chain fatty acids and their metabolic derivatives such as -hydroxy fatty acids and fatty acyl-CoA and acyl-L-carnitine esters. These substances might be detrimental for proper myocardial function. Previously, it has been suggested that intracellular lipid binding proteins like cytoplasmic fatty acid-binding protein (FABP) and acyl-CoA binding protein (ACBP) may bind these accumulating fatty acyl moieties to prevent their elevated levels from potentially harmful actions. In addition, the suggestion has been made that the abundantly present FABP may scavenge free radicals which are generated during reperfusion of the ischemic heart. However, these protective actions are challenged by the continuous physico-chemical partition of fatty acyl moieties between FABP and membrane structures and by the rapid release of FABP from ischemic and reperfused cardiac muscle. Careful evaluation of the available literature data reveals that at present no definite conclusion can be drawn about the potential protective effect of FABP on the ischemic and reperfused heart. Biochem123: 167–173, 1993)Abbreviations FABP
Fatty Acid-Binding Protein
- ACBP
Acyl-CoA Binding Protein
- MDGI
Mammary-Derived Growth Inhibitor
- CK
Creatine Kinase
- LDH
Lactate Dehydrogenase 相似文献
10.
A. Garnier C. Poizat C. Keriel P. Cuchet M. M. Vork Y. F. de Jong J. F. C. Glatz 《Molecular and cellular biochemistry》1993,123(1-2):107-112
The aim of this work was to study in the adult rat heart the effect of modifications of fatty acid (FA) supply on the content of cytoplasmic fatty acid-binding protein (H-FABPc). To modify the amount of circulating lipids, three different treatments were chosen: (i) an hypolipidemic treatment with Clofibrate, administered daily through a gastric tube at a dose of 250 mg/kg per day for one week, (ii) a continuous intravenous infusion of 20% Intralipid, a fat emulsion, for one week at a dose of 96 ml/kg per day, and (iii) a normobaric hypoxia exposure (pO2=10%) for three weeks. At the end of each treatment plasma lipids, myocardial H-FABPc content and the activities of three key enzymes (citrate synthase, CS, fructrose-6-phosphate kinase, FPK and hydroxy-acyl CoA-dehydrogenase, HAD) were assessed. With each of the three treatments a decrease of plasma cholesterol and phospholipid levels was observed. Plasma FA concentration increased with Intralipid infusion and decreased with chronic hypoxia. The heart H-FABPc content was increased by 20% with Clofibrate, decreased by 20% with chronic hypoxia and remained unaltered upon Intralipid treatment. The induced changes in H-FABPc content were not related directly to changes in plasma lipid levels. CS activity was slightly decreased in the hypoxia group, FPK activity decreased in the Clofibrate group, and HAD activity decreased in the Intralipid group. Among the various groups heart H-FABPc content was related to HAD activity. In conclusion, the H-FABPc content of adult rat heart appears responsive to changes in plasma lipid levels. 相似文献
11.
Members of the family of intracellular lipid binding proteins (iLBPs) have been implicated in cytoplasmic transport of lipophilic ligands, such as long-chain fatty acids and retinoids. iLBPs are low molecular mass proteins (14–16 kDa) sharing a common structural fold. The iLBP family likely arose through duplication and diversification of an ancestral iLBP gene. Phylogenetic analysis undertaken in the present study indicates that the ancestral iLBP gene arose after divergence of animals from fungi and plants. The first gene duplication was dated around 930 millions of years ago, and subsequent duplications in the succeeding 550 millions of years gave rise to the 16 iLBP types currently recognized in vertebrates. Four clusters of proteins, each binding a characteristic range of ligands, are evident from the phylogenetic tree. Evolution of different binding properties probably allowed cytoplasmic trafficking of distinct ligands. It is speculated that recruitment of an iLBP during evolution of animals enabled the mitochondrial oxidation of long-chain fatty acids. 相似文献
12.
Wolfgang Stremmel Hans-Erich Diede Enrique Rodilla-Sala Karel Vyska Monika Schrader Barbara Fitscher Salvatore Passarella 《Molecular and cellular biochemistry》1990,98(1-2):191-199
Summary For evaluation whether the membrane fatty acid-binding protein is related to mGOT, studies on the structure and function of both purified proteins were performed. Physicochemical characterization revealed that both proteins are different: the membrane fatty acid-binding protein has a molecular weight of 40 kD and a pI of 8.5–9.0, whereas rat mGOT has a molecular weight of 44 kD and a pI of 9.5–10.0. According to this distinct differences, they migrated separately on 2-dimensional electrophoresis. Furthermore, monospecific antibodies against the membrane fatty acid binding protein did not react with rat mGOT. In co-chromatography studies only the membrane fatty acid-binding protein showed affinity for long chain fatty acids, but not mGOT. Moreover, membrane binding studies were performed with the monospecific antibody to the membrane fatty acid binding protein. The inhibitory effect of this antibody on plasma membrane binding of oleate was reversed after preabsorption of the antibody with the membrane fatty acid binding protein, but was not affected after preabsorption with mGOT. These results indicate that the membrane fatty acid binding protein and mGOT are structurally and functionally not related. The data also support the significance of this membrane protein in the plasma membrane binding process of long chain fatty acids. 相似文献
13.
Fatty acid transport protein 1 and long-chain acyl coenzyme A synthetase 1 interact in adipocytes 总被引:1,自引:0,他引:1
The fatty acid transport proteins (FATP) and long-chain acyl coenzyme A synthetase (ACSL) proteins have been shown to play a role in facilitating long-chain fatty acid (LCFA) transport in mammalian cells under physiologic conditions. The involvement of both FATP and ACSL proteins is consistent with the model of vectorial acylation, in which fatty acid transport is coupled to esterification. This study was undertaken to determine whether the functions of these proteins are coordinated through a protein-protein interaction that might serve as a point of regulation for cellular fatty acid transport. We demonstrate for the first time that FATP1 and ACSL1 coimmunoprecipitate in 3T3-L1 adipocytes, indicating that these proteins form an oligomeric complex. The efficiency of FATP1 and ACSL1 coimmunoprecipitation is unaltered by acute insulin treatment, which stimulates fatty acid uptake, or by treatment with isoproterenol, which decreases fatty acid uptake and stimulates lipolysis. Moreover, inhibition of ACSL1 activity in adipocytes impairs fatty acid uptake, suggesting that esterification is essential for fatty acid transport. Together, our findings suggest that a constitutive interaction between FATP1 and ACSL1 contributes to the efficient cellular uptake of LCFAs in adipocytes through vectorial acylation. 相似文献
14.
Michael M. Vork Jan F. C. Glatz Don A. M. Surtel Ger J. van der Vusse 《Molecular and cellular biochemistry》1990,98(1-2):111-117
Summary Fatty acid (FA) binding by fatty acid-binding protein (FABP) is frequently Monitored with the so-called Lipidex 1000 assay, in which protein associated and non-protein bound FA are separated by selectively binding the latter to Lipidex 1000. Careful evaluation of this assay showed that the use of aqueous FA solutions resulted in a Marked decrease (60 to 70%) of FA concentration due to their aspecific binding to the surface of the test-tube used. In addition, solutions of rat heart FABP in the Molar range also showed a concentration decrease up to 80% due to protein binding to the surface of the test-tube. Introduction of detergents, Triton X-100 or Tween 20, limited the FA loss to less than 20% and totally eliminated FABP adsorption. Kinetic parameters for the binding of [1-14C]oleic acid by purified rat heart FABP, assayed in the presence of Triton X-100, were found to be similar to those assayed in the absence of detergent, when adequate corrections were Made for losses of FA and FABP due to surface adsorption. Use of Tween 20 resulted in a substantial increase of the dissociation constant. The addition of 100 M Triton X-100 to the assay medium considerably facilitates the determination of kinetic parameters of fatty acid-binding by proteins. 相似文献
15.
Torsten Börchers Peter Højrup Søren U. Nielsen Peter Roepstorff Friedrich Spener Jens Knudsen 《Molecular and cellular biochemistry》1990,98(1-2):127-133
Summary Cardiac-type fatty acid-binding protein (cFABP) from human heart muscle of three individuals was isolated and characterized as pI 5.3-cFABP. The proteins were structurally analyzed by tryptic peptide mapping, application of plasma desorption time-of-flight mass spectrometry and amino acid sequencing. All three preparations of human heart FABP, having 132 amino acids, differed from the published sequence [Offner et al. Biochem J 251: 191–198, 1988] in position 104, where Leu is found instead of Lys, and in position 124, where Cys is found instead of Ser. 相似文献
16.
Apple H. Kleine Jan F. C. Glatz Frans A. van Nieuwenhoven Monique I. J. Vallingal Martin H. L Salden Fré T. Bosman Wim J. A. Boersma Netty D. Zegers Ger J. van der Vusse 《Molecular and cellular biochemistry》1990,98(1-2):41-48
Summary In order to develop specific antibodies against human heart cytoplasmic fatty acid-binding protein (HFABPc), four oligo-peptides of 15–20 amino-acids each and corresponding with different antigenic parts of the human H-FABPc molecule, were synthesized. Polyclonal antibodies against these synthetic peptides were raised in mice (Balb/C) and rabbits (Flemish giant). When tested in enzyme linked immunosorbent assays (ELISA, antibody-capture assay), antisera against three of the four peptides showed a high immunoreactivity with the synthetic peptide selected for immunization as well as with the native human H-FABPc. Some cross-reactivity with the other synthetic peptides was observed for the rabbit antisera but not for those from mice. Polyclonal antibodies against synthetic peptides can be applied for the specific detection of the native protein in biological preparations containing proteins that show a high degree of homology with the protein to be assayed. 相似文献
17.
Soren U. Nielsen Anders O. Pedersen Henrik Vorum Rolf Brodersen 《Molecular and cellular biochemistry》1990,98(1-2):119-125
Summary A group of low molecular weight fatty acid-binding cytosolic proteins, FABPc with high abundance in heart, liver, skeletal muscle, intestine and adipose tissue, are anticipated to play a role in long-chain fatty acid metabolism in these tissues. Recently, a FABPc with MT 15 kDa has been purified from human heart muscle and found to be present in levels 2–4% of cytosolic proteins of human heart myocytes. In the present study two-dimensional gel electrophoresis under native and denaturing conditions has been used to characterize FABPc from human heart and this protein is found to be a major protein of human heart myocytes. The pI of FABPc from human heart was found to be about 5.3 under native conditions and about 6.5 in the presence of 9 M urea. 相似文献
18.
Ellen A. Meijer Sacco C. de Vries Peter Sterk Dorus W. J. Gadella Jr. Karel W. A. Wirtz Theo Hendriks 《Molecular and cellular biochemistry》1993,123(1-2):159-166
The extracellular protein EP2 was previously identified as non-specific lipid transfer protein based on its cDNA-derived amino acid sequence. Here, the purification of the EP2 protein from the medium of somatic embryo cultures is described. After two cycles of ion-exchange and gel permeation chromatography, a single silver-stained protein band with an apparent molecular mass of 10 kDa was observed on SDS-PAGE. This protein band was recognized by the antiserum raised against a EP2--galactosidase fusion-protein. Employing a fluorescent phospholipid analog, it was shown that the purified EP2 protein is capable of binding phospholipids and is able to enhance their transfer between artificial membranes. Employing a gel permeation assay, it could be demonstrated that the EP2 protein is also capable of binding palmitic and oleic acid as well as oleyl-CoA. Because in plants these fatty acids are used as precursor molecules for cutin, these results are in support of the proposed role of the EP2 protein to transport cutin monomers from their site of synthesis through the cell wall of epidermal cells to sites of cutin polymerization. 相似文献
19.
20.
Sheng-Li Zhou Barry J. Potter Decherd Stump Dario Sorrentino Paul D. Berk 《Molecular and cellular biochemistry》1990,98(1-2):183-189
Summary A plasma membrane fatty acid-binding protein (h-FABPPm) has been isolated from rat hepatocytes. Analogous proteins have also been identified in adipocytes, jejunal enterocytes and cardiac myocytes, all cells with high transmembrane fluxes of fatty acids. These 43 kDa, highly basic (pl = 9.1) FABPpm 's appear unrelated to the smaller, cytosolic FABP's (designated FABP's) identified previously in the same tissues. h-FABPpm appears closely related to the mitochondrial isoform of glutamic-oxaloacetic transaminase (mGOT), and both the purified protein and liver cell plasma membranes (LPM) possess GOT enzymatic activity. From their relative GOT specific activities it is estimated that h-FABPpm constitutes approximately 2% of LPM protein, or about 0.7 × 107 sites per cell. A monoclonal antibody-based competitive inhibition enzyme immunoassay (CIEIA) for h-FABPpm is described; it yields an estimate of 3.4 x 107 h-FABPpm sites per hepatocyte. Quantitated by either method, h-FABPPm appears to be a highly abundant protein constituent of LPM. 相似文献