Secretion of a novel, developmentally regulated fatty acid-binding protein into the perivitelline fluid of the parasitic nematode, Ascaris suum.
AuthorsMei B, Kennedy MW, Beauchamp J, Komuniecki PR, Komuniecki R
JournalJ Biol Chem
PubMed ID9092532
'Early development of the parasitic nematode, Ascaris suum, occurs inside a highly resistant eggshell, and the developing larva is bathed in perivitelline fluid. Two-dimensional gel analysis of perivitelline fluid from infective larvae reveals seven major proteins; a cDNA encoding one of these, As-p18, has been cloned, sequenced, and protein expressed ... More
Groups with polar characteristics can locate at both shallow and deep locations in membranes: the behavior of dansyl and related probes.
AuthorsAsuncion-Punzalan E, Kachel K, London E
JournalBiochemistry
PubMed ID9521780
'To understand the relationship between the chemical structure of polar molecules and their membrane location, the behavior of dansyl (dimethylaminonaphthalenesulfonyl) and related polar fluorescent probes was examined. The depth of these probes in lipid bilayers was determined by parallax analysis of fluorescence quenching [Chattopadhyay and London (1987) Biochemistry 26, 39-45; ... More
A novel lipid-binding protein from the cestode Moniezia expansa.
AuthorsJanssen D, Barrett J
JournalBiochem J
PubMed ID7575480
'A lipid-binding protein (LBP) has been purified from the cytosol of the cestode Moniezia expansa. The native LBP was found to be an oligomer of approx. 250 kDa, consisting of 11 kDa monomers. The LBP bound saturated and unsaturated fatty acids, but not their CoA derivatives, with KD values in ... More
High-performance liquid chromatographic determination of fatty acid binding proteins in rat liver with fluorescence detection.
AuthorsYamaguchi M, Wada K, Ishida J, Nakamura M
JournalAnalyst
PubMed ID1285576
'A highly sensitive, simple and reproducible method for the quantitative determination of fatty acid binding protein in rat liver by post-column high-performance liquid chromatography with fluorescence detection is presented. Fatty acid binding protein in rat liver cytosols is separated by gel-permeation column chromatography, which is followed by fluorescence detection. The ... More
Binding studies of tear lipocalin: the role of the conserved tryptophan in maintaining structure, stability and ligand affinity.
'The principal lipid binding protein in tears, tear lipocalin (TL), binds acid and the fluorescent fatty acid analogs, DAUDA and 16-AP at one site TL compete for this binding site. A fluorescent competitive binding assay revealed that apo-TL has a high affinity for phospholipids and stearic acid (Ki) of 1.2 ... More
A fluorometric determination of sphingomyelinase by use of fluorescent derivatives of sphingomyelin, and its application to diagnosis of Niemann-Pick disease.
AuthorsGatt S, Dinur T, Barenholz Y
JournalClin Chem
PubMed ID6243257
'We synthesized fluorescent derivatives of sphingomyelin (N-acyl-sphingosylphosphocholine) and used them as substrates for several sphingomyelinases. The following five fluorescent probes, each attached to the terminal carbon atom of the fatty acyl residue, were introduced into sphingomyelin: dansyl, pyrene, carbazole, 4-chloro-7-nitrobenz-2-oxa-1,3-diazole, and anthroic acid. We compared the rates at which the ... More
Solvent relaxation in lipid bilayers with dansyl probes.
AuthorsStubbs CD, Meech SR, Lee AG, Phillips D
JournalBiochim Biophys Acta
PubMed ID3995032
'The solvent relaxation properties of the dansyl group attached to two lipids (dansylphosphatidylethanolamine and dansylphosphatidylserine), a fatty acid (dansylundecanoic acid), and two drugs (dansylbenzocaine and dansylpropranolol) were compared in a variety of different lipid systems. Several methods for characterising solvent relaxation were compared in detail for dansylpropranolol in bilayer vesicles ... More
Gene structure and M20T polymorphism of the Schistosoma mansoni Sm14 fatty acid-binding protein. Molecular, functioanl, and immunoprotection analysis.
AuthorsRamos CR, Figueredo RC, Pertinhez TA, Vilar MM, do Nascimento AL, Tendler M, Raw I, Spisni A, Ho PL
JournalJ Biol Chem
PubMed ID12551912
'The Schistosoma mansoni Sm14 antigen belongs to the fatty acid-binding protein family and is considered a vaccine candidate against at least two parasite worms, Fasciola hepatica and S. mansoni. Here the genomic sequence and the polymorphism of Sm14 have been characterized for the first time. We found that the conserved ... More
Hydrophobic ligand binding by Zn-alpha 2-glycoprotein, a soluble fat-depleting factor related to major histocompatibility complex proteins.
AuthorsKennedy MW, Heikema AP, Cooper A, Bjorkman PJ, Sanchez LM
JournalJ Biol Chem
PubMed ID11425849
'Zn-alpha(2)-glycoprotein (ZAG) is a member of the major histocompatibility complex (MHC) class I family of proteins and is identical in amino acid sequence to a tumor-derived lipid-mobilizing factor associated with cachexia in cancer patients. ZAG is present in plasma and other body fluids, and its natural function, like leptin''s, probably ... More
Formation of membrane domains during the activation of protein kinase C.
AuthorsYang L, Glaser M
JournalBiochemistry
PubMed ID8909294
'The lateral membrane organization of phosphatidylserine, diacylglycerol, substrate, and Ca(2+)-dependent protein kinase C in large unilamellar vesicles was investigated by using fluorescence digital imaging microscopy. The formation of phosphatidylserine domains could be induced by either Ca2+, the MARCKS peptide, or protein kinase C. However, only Ca2+ could induce diacylglycerol to ... More
On the binding preference of human groups IIA and X phospholipases A2 for membranes with anionic phospholipids.
'Mammals contain 9-10 secreted phospholipases A(2) (sPLA(2)s) that display widely different affinities for membranes, depending on the phospholipid composition. The much higher enzymatic activity of human group X sPLA(2) (hGX) compared with human group IIA sPLA(2) (hGIIA) on phosphatidylcholine (PC)-rich vesicles is due in large part to the higher affinity ... More
Site-directed tryptophan fluorescence reveals the solution structure of tear lipocalin: evidence for features that confer promiscuity in ligand binding.
'The solution structure of human TL was deduced from the position of the emission peaks after site-directed tryptophan fluorescence (SDTF). The fluorescent amino acid tryptophan was sequentially substituted for each native amino acid in the sequence. Characteristic periodicities for eight beta-strands that comprise the beta-barrel and three alpha-helices were identified. ... More
Bacterial cell membrane hydrolysis by secreted phospholipases A(2): a major physiological role of human group IIa sPLA(2) involving both bacterial cell wall penetration and interfacial catalysis.
AuthorsBuckland AG, Heeley EL, Wilton DC
JournalBiochim Biophys Acta
PubMed ID10760469
'The ability of human group IIa secreted phospholipase A(2) (human sPLA(2)) to hydrolyse the phospholipid membrane of whole cell suspensions of Gram-positive bacteria is demonstrated in real time using a continuous fluorescence displacement assay. Micrococcus luteus is used as a model system and demonstrates an almost absolute specificity for this ... More
The binding of natural and fluorescent lysophospholipids to wild-type and mutant rat liver fatty acid-binding protein and albumin.
AuthorsThumser AE, Wilton DC
JournalBiochem J
PubMed ID7717990
'Rat liver fatty acid-binding protein (FABP) is able to bind a wide range of non-polar anionic ligands, including lysophospholipids. In order to understand the nature of lysophospholipid interactions with liver FABP, the binding of natural lysophospholipids and two fluorescent analogues, N-(5-dimethylaminonaphthalenesulphonyl)-1-palmitoyl-sn-glycero-3- phosphoethanolamine (dansyl lysoPE) and 1-(O-[11-(5-dimethylaminonaphthalene-sulphonyl)amino]undecyl)-sn-glycero -3- phosphocholine (dansyl-C11-lysoPAF), has ... More
Comparison of the catalytic properties of phospholipase A2 from pancreas and venom using a continuous fluorescence displacement assay.
AuthorsKinkaid A, Wilton DC
JournalBiochem J
PubMed ID1898370
'Phospholipases A2 from pig pancreas and the venoms from bee, Naja naja and Crotalus atrox have been studied by using a new continuous fluorescence displacement assay that utilizes normal phospholipid substrates [Wilton (1990) Biochem. J. 266, 435-439]. With limiting amounts of substrate, the assay demonstrated stoichiometric conversion into products with ... More
Flexible loop of beta 2-glycoprotein I domain V specifically interacts with hydrophobic ligands.
AuthorsHong DP, Hagihara Y, Kato H, Goto Y
JournalBiochemistry
PubMed ID11434778
'Beta2-glycoprotein I (beta2-GPI), which consists of four complement control protein modules and a distinctly folded fifth C-terminal domain, is an essential cofactor for the binding to phospholipids of anti-cardiolipin antibodies, isolated from patients with anti-phospholipid antibody syndrome, and its fifth domain has attracted attention as a specific phospholipid-binding site. We ... More
A surface-associated retinol- and fatty acid-binding protein (Gp-FAR-1) from the potato cyst nematode Globodera pallida: lipid binding activities, structural analysis and expression pattern.
AuthorsPrior A, Jones JT, Blok VC, Beauchamp J, McDermott L, Cooper A, Kennedy MW
JournalBiochem J
PubMed ID11368765
'Parasitic nematodes produce at least two structurally novel classes of small helix-rich retinol- and fatty-acid-binding proteins that have no counterparts in their plant or animal hosts and thus represent potential targets for new nematicides. Here we describe a protein (Gp-FAR-1) from the plant-parasitic nematode Globodera pallida, which is a member ... More
Bacterial and host interactions during the biogenesis, toxicity and immunogenicity of Escherichia coli heat-labile enterotoxin.
Binding of amphipathic drugs and probes to biological membranes.
AuthorsMoules IK, Rooney EK, Lee AG
JournalFEBS Lett
PubMed ID6121718
Fitting fluorescence emission spectra of probes bound to biological membranes.
AuthorsRooney EK, Lee AG
JournalJ Biochem Biophys Methods
PubMed ID3711585
We show that fluorescence emission spectra for molecules containing the dansyl fluorophor can be accurately described as skewed Gaussians, and that spectra for dansyl probes bound to biological membranes can be resolved using least-squares techniques into two components, representing probe bound to the lipid and protein sites in the membrane. ... More
Synthesis of a gene for rat liver fatty-acid-binding protein and its expression in Escherichia coli.
AuthorsWorrall AF, Evans C, Wilton DC
JournalBiochem J
PubMed ID1898328
A gene coding for rat liver fatty-acid-binding protein (FABP) has been constructed by the single-step ligation of ten synthetic oligonucleotides. The gene has been cloned into the bacterial expression vector pKK223-3. Induction of protein synthesis from this gene results in over expression of an FABP that is indistinguishable in its ... More
Fluorescent probes for retinoic acid receptors: molecular measures for the ligand binding pocket.
AuthorsShimazawa R, Hibino S, Mizoguchi H, Hashimoto Y, Iwasaki S, Kagechika H, Shudo K
JournalBiochem Biophys Res Commun
PubMed ID1656968
Two fluorescent probes for nuclear retinoic acid receptors (RARs) have been developed, both containing a biologically active retinoid moiety and a fluorescent dansyl moiety, but differing in the length of the spacer arm connecting the two moieties. Both probes bind RARs at their retinoid-binding sites, revealing the usefulness of the ... More
A fluorescence displacement assay for the measurement of arachidonoyl ethanolamide (anandamide) and oleoyl amide (octadecenoamide) hydrolysis.
AuthorsThumser AE, Voysey J, Wilton DC
JournalBiochem Pharmacol
PubMed ID9065749
We describe a simple fluorescence displacement assay to measure hydrolysis of arachidonoyl ethanolamide and oleoyl amide, two important pharmacological compounds. Hydrolysis at the amide linkage of these ligands releases a fatty acid as one of the products. The displacement of a fluorescent fatty acid analogue from rat liver fatty acid-binding ... More
Studies on fatty acid-binding proteins. The detection and quantification of the protein from rat liver by using a fluorescent fatty acid analogue.
AuthorsWilkinson TC, Wilton DC
JournalBiochem J
PubMed ID3800946
Fatty acid-binding protein from rat liver is shown to bind the fluorescent fatty acid probe dansyl undecanoic acid. Binding is accompanied by a shift in the fluorescence emission maximum from 550 nm to 500 nm and a 60-fold fluorescence enhancement at 500 nm. These spectral properties have allowed the use ... More
Interaction of fatty acids with the calcium-magnesium ion dependent adenosinetriphosphatase from sarcoplasmic reticulum.
AuthorsLee AG, East JM, Jones OT, McWhirter J, Rooney EK, Simmonds AC
JournalBiochemistry
PubMed ID6129894
The fluorescence emission spectrum of dansylundecanoic acid is sensitive to the environment and appears at a lower wavelength when the fatty acid is bound to protein than when it is bound to phospholipid. When bound to the (Ca2+-Mg2+)-ATPase of sarcoplasmic reticulum, the emission spectrum can be resolved into separate components ... More
The binding of the fluorescent ATP analogue 2'(3')-trinitrophenyladenosine-5'-triphosphate to rat liver fatty acid-binding protein.
AuthorsSheridan M, Wilton DC
JournalFEBS Lett
PubMed ID1468590
The less polar fluorescent analogue of ATP, 2'(3')-trinitrophenyl-5'-triphosphate bound to rat liver fatty acid-binding protein with high affinity (Kd 6.3 x 10(-6) M) and 1:1 molar stoichiometry. This probe bound to the fatty acid binding site of the protein and was displaced by oleic acid and oleoyl CoA. High concentrations ... More
The fatty acid analogue 11-(dansylamino)undecanoic acid is a fluorescent probe for the bilirubin-binding sites of albumin and not for the high-affinity fatty acid-binding sites.
AuthorsWilton DC
JournalBiochem J
PubMed ID2396975
1. The fluorescent fatty acid probe 11-(dansylamino)undecanoic acid (DAUDA) binds with high affinity to bovine and human serum albumin (BSA and HSA) at three sites. 2. The Kd of the primary binding site could not be determined; however, the two secondary sites appeared to be equivalent, with an apparent Kd ... More
A continuous fluorescence displacement assay for the measurement of phospholipase A2 and other lipases that release long-chain fatty acids.
AuthorsWilton DC
JournalBiochem J
PubMed ID2317197
1. A new continuous fluorescence assay for phospholipase A2 is described which involves the displacement of the highly fluorescent fatty-acid probe 11-(dansylamino)undecanoic acid from rat liver fatty-acid-binding protein by long-chain fatty acids released as a result of phospholipase A2-catalysed hydrolysis of phospholipids. The initial rate of decrease in fluorescence is ... More
Time-resolved emission spectroscopy of the dansyl fluorescence probe.
AuthorsGhiggino KP, Lee AG, Meech SR, O'Connor DV, Phillips D
JournalBiochemistry
PubMed ID6895314
Using a cavity-dumped laser system as the light source, we have recorded time-resolved fluorescence spectra for the dansyl group in a variety of systems. The fluorescence spectrum of dansylphosphatidylethanolamine incorporated into a variety of lipid bilayer systems at ambient temperatures shows very marked time dependence, with red shifts of up ... More
The secretory Onchocerca volvulus protein OvS1/Ov20 exhibits the capacity to compete with serum albumin for the host's long-chain fatty acids.
AuthorsMpagi JL, Erttmann KD, Brattig NW
JournalMol Biochem Parasitol
PubMed ID10693749
The mechanism by which filarial parasites derive fatty acids bound to the host's carrier protein is poorly understood. The capacity of a secretory protein of Onchocerca volvulus (OvS1/Ov20) to compete with serum albumin for arachidonic and other fatty acids was investigated in this study. Binding affinities of the two proteins ... More
A continuous fluorescence displacement assay for phospholipase A2 using albumin and medium chain phospholipid substrates.
AuthorsKinkaid AR, Wilton DC
JournalAnal Biochem
PubMed ID8368517
A continuous fluorescence assay for phospholipase A2 is described which involves the displacement of the highly fluorescent fatty-acid probe 11-(dansylamino) undecanoic acid from albumin by decanoic acid released as a result of phospholipase A2-catalyzed hydrolysis of didecanoyl-phosphatidylcholine. Using the phospholipase A2 from Naja naja, this assay will detect activity from ... More
The crystal structure of the liver fatty acid-binding protein. A complex with two bound oleates.
AuthorsThompson J, Winter N, Terwey D, Bratt J, Banaszak L
JournalJ Biol Chem
PubMed ID9054409
The crystal structure of the recombinant form of rat liver fatty acid-binding protein was completed to 2.3 A and refined to an R factor of 19.0%. The structural solution was obtained by molecular replacement using superimposed polyalanine coordinates of six intracellular lipid-binding proteins as a search probe. The entire amino ... More
Interfacial kinetic and binding properties of the complete set of human and mouse groups I, II, V, X, and XII secreted phospholipases A2.
AuthorsSinger AG, Ghomashchi F, Le Calvez C, Bollinger J, Bezzine S, Rouault M, Sadilek M, Nguyen E, Lazdunski M, Lambeau G, Gelb MH
JournalJ Biol Chem
PubMed ID12359733
Expression of the full set of human and mouse groups I, II, V, X, and XII secreted phospholipases A(2) (sPLA(2)s) in Escherichia coli and insect cells has provided pure recombinant enzymes for detailed comparative interfacial kinetic and binding studies. The set of mammalian sPLA(2)s display dramatically different sensitivity to dithiothreitol. ... More
A continuous fluorescence-displacement assay for triacylglycerol lipase and phospholipase C that also allows the measurement of acylglycerols.
AuthorsWilton DC
JournalBiochem J
PubMed ID1903930
A new continuous fluorescence-displacement assay for enzymes that release long-chain fatty acids [Wilton (1990) Biochem. J. 266, 435-439] is described in detail for pig pancreatic triacylglycerol lipase. The assay involves the displacement of the highly fluorescent fatty acid probe 11-(dansylamino)undecanoic acid from rat liver fatty acid-binding protein by long-chain fatty ... More
Mutagenic and chemical modification of the ABA-1 allergen of the nematode Ascaris: consequences for structure and lipid binding properties.
AuthorsMcDermott L, Moore J, Brass A, Price NC, Kelly SM, Cooper A, Kennedy MW
JournalBiochemistry
PubMed ID11502186
The polyprotein allergens/antigens of nematodes (NPAs) are the only lipid binding proteins known to be produced as polyproteins. Cleavage of the large polyprotein precursors at regularly spaced proteinase cleavage sites produces 10 or 11 individual protein units of approximately 15 kDa. The sequences of these units are highly diverse within ... More
The Ov20 protein of the parasitic nematode Onchocerca volvulus. A structurally novel class of small helix-rich retinol-binding proteins.
AuthorsKennedy MW, Garside LH, Goodrick LE, McDermott L, Brass A, Price NC, Kelly SM, Cooper A, Bradley JE
JournalJ Biol Chem
PubMed ID9368002
Ov20 is a major antigen of the parasitic nematode Onchocerca volvulus, the causative agent of river blindness in humans, and the protein is secreted into the tissue occupied by the parasite. DNA encoding Ov20 was isolated, and the protein was expressed in Escherichia coli. Fluorescence-based ligand binding assays show that ... More
Formation of membrane domains by the envelope proteins of vesicular stomatitis virus.
AuthorsLuan P, Glaser M
JournalBiochemistry
PubMed ID8161502
The properties of the two envelope-associated proteins of vesicular stomatitis virus, the glycoprotein (G) and the matrix protein (M), were investigated in order to understand the mechanism of virus budding and domain formation in membranes. Fluorescence resonance energy transfer was used to study the interaction between the G protein and ... More
Effects of fluorophore structure and hydrophobicity on the uptake and metabolism of fluorescent lipid analogs.
AuthorsNaylor BL, Picardo M, Homan R, Pownall HJ
JournalChem Phys Lipids
PubMed ID1934193
Cellular transport and metabolism of fatty acids are integral components of lipid metabolism, but the mechanisms and regulation involved are poorly understood. A variety of commercially available fluorescent analogs of fatty acids, are potentially useful probes for the study of lipid metabolism by such techniques as cell sorting and fluorescence ... More
The DvA-1 polyprotein of the parasitic nematode Dictyocaulus viviparus. A small helix-rich lipid-binding protein.
AuthorsKennedy MW, Britton C, Price NC, Kelly SM, Cooper A
JournalJ Biol Chem
PubMed ID7642601
DNA encoding a single unit of the DvA-1 polyprotein of the parasitic nematode Dictyocaulus viviparus was isolated and the polypeptide ("rDvA-1L") expressed in Escherichia coli, to give a protein showing high binding affinity for fatty acids and retinoids. Fluorescent fatty acid probes show substantial changes in emission spectrum in the ... More
The binding of lysophospholipids to rat liver fatty acid-binding protein and albumin.
AuthorsThumser AE, Voysey JE, Wilton DC
JournalBiochem J
PubMed ID8053904
The binding of lysophospholipids to rat liver fatty acid-binding protein (FABP) and to BSA and human serum albumin was investigated by using competitive displacement fluorescence assays by monitoring the displacement of the fluorescent fatty acid probe 11-(dansylamino)undecanoic acid (DAUDA). In addition, direct binding assays using changes in tryptophan fluorescence were ... More
Serum constituents can affect 2'-& 3'-O-(4-benzoylbenzoyl)-ATP potency at P2X(7) receptors.
AuthorsMichel AD, Xing M, Humphrey PP
JournalBr J Pharmacol
PubMed ID11264244
1. 2'-& 3'-O-(4-benzoylbenzoyl)-ATP (BzATP) is the prototypic agonist for P2X(7) receptors. In this study we demonstrate that bovine serum albumin (BSA) can affect the potency of BzATP at P2X receptors. 2. BzATP potency (pEC(50)) to stimulate ethidium accumulation in cells expressing recombinant P2X7 receptors varied between 6.5 and 4, depending ... More
Ammodytoxins, potent presynaptic neurotoxins, are also highly efficient phospholipase A2 enzymes.
AuthorsPetan T, Krizaj I, Gelb MH, Pungercar J
JournalBiochemistry
PubMed ID16156665
The enzymatic activity of ammodytoxins (Atxs), secreted phospholipases A(2) (sPLA(2)s) in snake venom, is essential for expression of their presynaptic neurotoxicity, but its exact role in the process is unknown. We have analyzed in detail the enzymatic properties of Atxs, their mutants, and homologues. The apparent rates of phospholipid hydrolysis ... More
Zn-alpha2-glycoprotein, an MHC class I-related glycoprotein regulator of adipose tissues: modification or abrogation of ligand binding by site-directed mutagenesis.
AuthorsMcDermott LC, Freel JA, West AP, Bjorkman PJ, Kennedy MW
JournalBiochemistry
PubMed ID16475792
Zn-alpha(2)-glycoprotein (ZAG) is a soluble lipid-mobilizing factor associated with cancer cachexia and is a novel adipokine. Its X-ray crystal structure reveals a poly(ethylene glycol) molecule, presumably substituting for a higher affinity natural ligand, occupying an apolar groove between its alpha(1) and alpha(2) domain helices that corresponds to the peptide binding ... More
Distribution of phospholipids around gramicidin and D-beta-hydroxybutyrate dehydrogenase as measured by resonance energy transfer.
AuthorsWang S, Martin E, Cimino J, Omann G, Glaser M
JournalBiochemistry
PubMed ID2454133
A resonance energy transfer method was developed to study the distribution of phospholipids around integral membrane proteins. The method involved measuring the extent of energy transfer from tryptophan residues of the proteins to different phospholipids labeled with a dansyl moiety in the fatty acid chain. No specific interactions were observed ... More
Visualization of Ca2+-induced phospholipid domains.
AuthorsHaverstick DM, Glaser M
JournalProc Natl Acad Sci U S A
PubMed ID3474616
Large vesicles (5-15 microns) were formed by hydrating a dried lipid film containing phospholipids labeled with a fluorophore in one fatty acid chain. By using a fluorescence microscope attached to a low-light-intensity charge-coupled-device camera and digital-image processor, the vesicles were easily viewed and initially showed uniform fluorescence intensity across the ... More
Combined effects of sphingomyelin and cholesterol on the hydrolysis of emulsion particle triolein by lipoprotein lipase.
AuthorsLobo LI, Wilton DC
JournalBiochim Biophys Acta
PubMed ID9421185
Sphingomyelin (SM) is one of the major lipids in lipoproteins. However, its function in lipoprotein metabolism is unknown. In an attempt to understand the role that this lipid plays in modulation of lipoprotein lipase (LPL)-mediated hydrolysis, triolein-based emulsion particles containing 15% (physiological concentration) and 30% of the phospholipid content as ... More
The ABA-1 allergen of the parasitic nematode Ascaris suum: fatty acid and retinoid binding function and structural characterization.
AuthorsKennedy MW, Brass A, McCruden AB, Price NC, Kelly SM, Cooper A
JournalBiochemistry
PubMed ID7756301
We report here on the structure and function of the ABA-1 allergen protein of the parasitic nematode Ascaris, the first nematode allergen to be characterized in detail. Using the fluorescent fatty acid analog 11-(((5-(dimethylamino)-1-naphthalenyl)sulfonyl)amino)undecanoic acid (DAUDA), it was demonstrated that ABA-1 is a fatty acid binding protein (FABP) with a ... More
Tryptophan-containing mutant of human (group IIa) secreted phospholipase A2 has a dramatically increased ability to hydrolyze phosphatidylcholine vesicles and cell membranes.
AuthorsBaker SF, Othman R, Wilton DC
JournalBiochemistry
PubMed ID9748327
Human nonpancreatic (group IIa) secreted phospholipase A2 (human sPLA2) is associated with a number of inflammatory disorders in which the extracellular concentrations of this enzyme can become highly elevated. It is probable that the enzyme normally acts as an acute-phase protein whose function is to facilitate the removal of infectious ... More
The FAR protein family of the nematode Caenorhabditis elegans. Differential lipid binding properties, structural characteristics, and developmental regulation.
AuthorsGarofalo A, Rowlinson MC, Amambua NA, Hughes JM, Kelly SM, Price NC, Cooper A, Watson DG, Kennedy MW, Bradley JE
JournalJ Biol Chem
PubMed ID12502713
Parasitic nematodes of humans and plants secrete a structurally novel type of fatty acid- and retinol-binding protein, FAR, into the tissues they occupy. These proteins may interfere with intercellular lipid signaling to manipulate the defense reactions of the host or acquire essential lipids for the parasites. The genome of the ... More
Expression of human liver fatty acid-binding protein in Escherichia coli and comparative analysis of its binding characteristics with muscle fatty acid-binding protein.
AuthorsMaatman RG, van Moerkerk HT, Nooren IM, van Zoelen EJ, Veerkamp JH
JournalBiochim Biophys Acta
PubMed ID8068722
Human liver fatty acid-binding protein (L-FABP) has been efficiently expressed in Escherichia coli. The cDNA encoding human liver FABP was under the control of T7 RNA polymerase promoter in the expression vector pET-3b. Expression required overnight induction with isopropyl beta-D-thiogalactopyranoside in the presence of the bacterial RNA polymerase inhibitor, rifampicin. ... More
Binding of recombinant rat liver fatty acid-binding protein to small anionic phospholipid vesicles results in ligand release: a model for interfacial binding and fatty acid targeting.
AuthorsDavies JK, Thumser AE, Wilton DC
JournalBiochemistry
PubMed ID10606528
A number of intracellular proteins bind to negatively charged phospholipid membranes, and this interfacial binding results in a conformational change that modulates the activity of the protein. Using a fluorescent fatty acid analogue, 11-[5-(dimethylamino)naphthalenesulfonyl]undecanoic acid (DAUDA), it is possible to demonstrate the release of this ligand from recombinant rat liver ... More
Effect of charge reversal mutations on the ligand- and membrane-binding properties of liver fatty acid-binding protein.
AuthorsDavies JK, Hagan RM, Wilton DC
JournalJ Biol Chem
PubMed ID12379651
Liver fatty acid-binding protein (FABP) is able to bind to anionic phospholipid vesicles under conditions of low ionic strength. This binding results in the release of ligand, the fluorescent fatty acid analogue 11-dansylaminoundecanoic acid (DAUDA), with loss of fluorescence intensity (Davies, J. K., Thumser, A. E. A., and Wilton, D. ... More