S100P, a novel Ca(2+)-binding protein from human placenta. cDNA cloning, recombinant protein expression and Ca2+ binding properties.
AuthorsBecker T, Gerke V, Kube E, Weber K
JournalEur J Biochem
PubMed ID1633809
A novel member of the S100 protein family, present in human placenta, has been characterized by protein sequencing, cDNA cloning, and analysis of Ca(2+)-binding properties. Since the placenta protein of 95 amino acid residues shares about 50% sequence identity with the brain S100 proteins alpha and beta, we proposed the ... More
Resolution of multicomponent fluorescence emission by phase sensitive detection at multiple modulation frequencies.
AuthorsKeating-Nakamoto SM, Cherek H, Lakowicz JR
JournalAnal Chem
PubMed ID3619043
Probing the interactions of alcohols with biological membranes with the fluorescent probe Prodan.
AuthorsRottenberg H
JournalBiochemistry
PubMed ID1390730
'Prodan [6-propionyl-2-(dimethylamine)naphthalene] is a hydrophobic fluorescent probe which is extremely sensitive to both the polarity and the hydrogen-bond donating capacity of the solvent. In binary mixtures of solvents, the hydrogen-bond donating effect on Prodan fluorescence saturates at relatively low concentrations of protic solvent while the polarity effect is proportional to ... More
Investigation of the lipid domains and apolipoprotein orientation in reconstituted high density lipoproteins by fluorescence and IR methods.
AuthorsWald JH, Coormaghtigh E, De Meutter J, Ruysschaert JM, Jonas A
JournalJ Biol Chem
PubMed ID2123199
'The reconstituted high density lipoproteins (rHDL) that were described in the preceding paper (Hefele Wald, J., Krul, E. S., and Jonas, A. (1990) J. Biol. Chem. 265, 20037-20043) are used in this study to analyze the organization, conformation, and dynamics of the lipid phase, as well as the relative orientation ... More
Dipolar solvent relaxation on a nanosecond time scale in ether phospholipid membranes as determined by multifrequency phase and modulation fluorometry.
AuthorsSommer A, Paltauf F, Hermetter A
JournalBiochemistry
PubMed ID2271701
'The present study reports on the observation of dipolar solvent relaxation in phospholipid membranes using multifrequency phase and modulation fluorometry. We measured the time-resolved emission spectra of 6-propionyl-2-(dimethylamino)naphthalene (PRODAN) in artificial bilayer membranes of chemically defined acyl-, alkyl-, and alkenyl-substituted phospholipids at 15 degrees C. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, 3-O-hexadecyl-2-oleoyl-sn-glycero-1-phosphocholine, or 1-O-hexadec-1''-enyl-2-oleoyl-sn-glycero-3-phosphocholine (plasmalogen) ... More
Thermotropic properties of phosphatidylethanols.
AuthorsBondar OP, Rowe ES
JournalBiophys J
PubMed ID8874018
'Phosphatidylethanol is formed when ethanol substitutes in the transphosphatidylation reaction catalyzed by phospholipase D. The structural and thermotropic properties of dipalmitoylphosphatidylethanol and dimyristoylphosphatidylethanol have been studied using differential scanning calorimetry, fluorescence spectroscopy, and 31P nuclear magnetic resonance. These lipids exist in a bilayer phase with no indication of nonbilayer phase ... More
Organization and dynamics of tryptophan residues in erythroid spectrin: novel structural features of denatured spectrin revealed by the wavelength-selective fluorescence approach.
AuthorsChattopadhyay A, Rawat SS, Kelkar DA, Ray S, Chakrabarti A
JournalProtein Sci
PubMed ID14573853
'We have investigated the organization and dynamics of the functionally important tryptophan residues of erythroid spectrin in native and denatured conditions utilizing the wavelength-selective fluorescence approach. We observed a red edge excitation shift (REES) of 4 nm for the tryptophans in the case of spectrin in its native state. This ... More
Interactions of Laurdan with phosphatidylcholine liposomes: a high pressure FTIR study.
AuthorsChong PL, Wong PT
JournalBiochim Biophys Acta
PubMed ID8323945
'The interactions of 6-lauroyl-2-dimethylaminonaphthalene (Laurdan) with L-alpha-dimyristoylphosphatidylcholine (DMPC) have been studied isothermally at 28 degrees C by Fourier-transform infrared spectroscopy (FTIR) at two pH values (6.8 and 3.0) and over the pressure range of 0.001-25 kbar. The results obtained with Laurdan are compared with those previously obtained with 6-propionyl-2-dimethylaminonaphthalene (Prodan) ... More
Interaction of myosin.ADP.fluorometal complexes with fluorescent probes and direct observation using quick-freeze deep-etch electron microscopy.
AuthorsMaruta S, Uyehara Y, Aihara T, Katayama E
JournalJ Biochem (Tokyo)
PubMed ID15269240
'Myosin forms stable ternary complexes with ADP and phosphate analogues of fluorometals that mimic different ATPase reaction intermediates corresponding to each step of the cross-bridge cycle. In the present study, we monitored the formation of ternary complexes of myosin.ADP.fluorometal using the fluorescence probe prodan. It has been reported that the ... More
The influence of cholesteryl ester transfer protein on the composition, size, and structure of spherical, reconstituted high density lipoproteins.
AuthorsRye KA, Hime NJ, Barter PJ
JournalJ Biol Chem
PubMed ID7814372
'The effect of cholesteryl ester transfer protein (CETP) on the size, composition, and structure of spherical, reconstituted HDL (rHDL) which contain apolipoprotein (apo) A-I as their sole apolipoprotein has been studied. Spherical rHDL were incubated with CETP and Intralipid for up to 24 h. During this time CETP promoted transfers ... More
Prodan as a membrane surface fluorescence probe: partitioning between water and phospholipid phases.
AuthorsKrasnowska EK, Gratton E, Parasassi T
JournalBiophys J
PubMed ID9545057
'Fluorescence spectral features of 6-propionyl-2-dimethylaminonaphthalene (Prodan) in phospholipid vesicles of different phase states are investigated. Like the spectra of 6-lauroyl-2-dimethylaminonaphthalene (Laurdan), the steady-state excitation and emission spectra of Prodan are sensitive to the polarity of the environment, showing a relevant shift due to the dipolar relaxation phenomenon. Because of the ... More
The interaction of 6-propionyl-2-(NN-dimethyl)aminonaphthalene (PRODAN)-labelled actin with actin-binding proteins and drugs.
AuthorsZechel K
JournalBiochem J
PubMed ID8452529
'The influence of various actin-binding proteins and drugs on the fluorescence emission of rabbit muscle actin labelled with the fluorescent probe acrylodan (6-acryloyl-2-dimethylaminonaphthalene) at Cys-374, the penultimate amino acid residue of the actin amino acid sequence, was studied. Addition of myosin, tropomyosin or phalloidin, agents known to bind only to ... More
Analysis of excited-state processes by phase-modulation fluorescence spectroscopy.
AuthorsLakowicz JR, Balter A
JournalBiophys Chem
PubMed ID7139044
'Fluorescence phase shift and demodulation methods were used in the analysis of excited-state reactions and to investigate solvent relaxation around fluorophores in viscous solvents. The chosen samples illustrate the results expected for fluorophores bound to biological macromolecules. These moderately simple samples served to test the theoretical predictions described in the ... More
Different modulation of phospholipase A2 activity by saturated and monounsaturated N-acylethanolamines.
AuthorsZolese G, Wozniak M, Mariani P, Saturni L, Bertoli E, Ambrosini A
JournalJ Lipid Res
PubMed ID12562826
'The physiological functions of N-acylethanolamines (NAEs) are poorly understood, although many functions were suggested for these naturally occurring membrane components of plants and animals. The binding with cannabinoid receptors CB1 and CB2 was demonstrated for some NAEs, such as anandamide. However, the chemical nature of these molecules suggests that some ... More
Phase-sensitive fluorescence spectroscopy: a new method to resolve fluorescence lifetimes or emission spectra of components in a mixture of fluorophores.
AuthorsLakowicz JR, Cherek H
JournalJ Biochem Biophys Methods
PubMed ID7276422
'A novel phase fluorometric method is described which permits direct recording of individual emission spectra from a mixture of two fluorescent compounds. Additionally, the lifetimes of each component may be determined by examination of the phase-sensitive fluorescence spectra. The method utilizes phase-sensitive detection of the sinusoidally modulated emission from a ... More
Binding of lecithin:cholesterol acyltransferase to reconstituted high density lipoproteins is affected by their lipid but not apolipoprotein composition.
AuthorsBolin DJ, Jonas A
JournalJ Biol Chem
PubMed ID8125962
'The reaction of lecithin:cholesterol acyltransferase (LCAT) with high density lipoproteins (HDL) is of critical importance in reverse cholesterol transport. We studied the relationship between LCAT binding and HDL composition using two assays to determine LCAT binding affinity for discoidal reconstituted HDL (rHDL). We prepared rHDL with egg phosphatidylcholine (egg-PC), cholesterol, ... More
The fluorescent probe prodan characterizes the warfarin binding site on human serum albumin.
AuthorsMoreno F, Cortijo M, González-Jiménez J
JournalPhotochem Photobiol
PubMed ID10063798
'The fluorescent probe Prodan (6-propionyl-2-dimethyl-aminonaphthalene) binds with high affinity to human serum albumin (HSA). The spectral characteristics of the Prodan bound to the protein are very different from the free Prodan in solution. These differences allowed the spectra to be deconvoluted into log-normal bands in order to quantify the bound ... More
Relationships between bilayer structure and phospholipase A2 activity: interactions among temperature, diacylglycerol, lysolecithin, palmitic acid, and dipalmitoylphosphatidylcholine.
AuthorsBell JD, Burnside M, Owen JA, Royall ML, Baker ML
JournalBiochemistry
PubMed ID8664287
'Bilayers composed of phosphatidylcholine initially resist catalysis by phospholipase A2. However, after a latency period, they become susceptible when sufficient reaction products (lysolecithin and fatty acid) accumulate in the membrane. Temperatures near the main bilayer phase transition and saturated long-chain diacylglycerol in the bilayer modulate the effectiveness of the reaction ... More
The influence of sphingomyelin on the structure and function of reconstituted high density lipoproteins.
AuthorsRye KA, Hime NJ, Barter PJ
JournalJ Biol Chem
PubMed ID8626769
'The effect of sphingomyelin (SPM) on the structure and function of discoidal and spherical reconstituted high density lipoproteins (rHDL) has been studied. Three preparations of discoidal rHDL with 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC)/SPM/unesterified cholesterol (UC)/apolipoprotein (apo)A-I molar ratios of 99.6/0. 0/10.2/1.0, 86.0/13.6/10.8/1.0, and 72.5/26.3/11.4/1.0 were prepared by cholate dialysis. SPM did not ... More
Effect of vesicle size on the prodan fluorescence in diheptadecanoylphosphatidylcholine bilayer membrane under atmospheric and high pressures.
AuthorsGoto M, Sawaguchi H, Tamai N, Matsuki H, Kaneshina S,
JournalLangmuir
PubMed ID20695581
'The bilayer phase behavior of diheptadecanoylphosphatidylcholine (C17PC) with different vesicle sizes (large multilamellar vesicle (LMV) and giant multilamellar vesicle (GMV)) was investigated by fluorescence spectroscopy using a polarity-sensitive fluorescent probe Prodan under atmospheric and high pressures. The difference in phase transitions and thermodynamic quantities of the transition was hardly observed ... More
Dipole potential and head-group spacing are determinants for the membrane partitioning of pregnanolone.
'The membrane interactions of pregnanolone, an intravenous general anesthetic steroid, were characterized using fluorescence spectroscopy and monolayer technique. di-8-ANEPPS [4-[2-[6-(dioctylamino)-2-naphthalenyl]ethenyl]-1-(3-sulfopropyl)-pyridinium], a membrane dipole potential (Psi)-sensitive probe, revealed pregnanolone to decrease Psi similarly as reported previously for other anesthetics. The decrement in Psi was approximately 16 and 10 mV in dipalmitoylphosphatidylcholine ... More
Surface properties of native human plasma lipoproteins and lipoprotein models.
AuthorsMassey JB, Pownall HJ
JournalBiophys J
PubMed ID9533698
'Plasma lipoprotein surface properties are important but poorly understood determinants of lipoprotein catabolism. To elucidate the relation between surface properties and surface reactivity, the physical properties of surface monolayers of native lipoproteins and lipoprotein models were investigated by fluorescent probes of surface lipid fluidity, surface lateral diffusion, and interfacial polarity, ... 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
Bimodal distribution and fluorescence response of environment-sensitive probes in lipid bilayers.
AuthorsKlymchenko AS, Duportail G, Demchenko AP, Mély Y
JournalBiophys J
PubMed ID15111409
'A remarkable heterogeneity is often observed in the spectroscopic properties of environment-sensitive fluorescence probes in phospholipid bilayers. To explain its origin, we provided a detailed investigation of the fluorescence excitation and emission spectra of 4''-dimethylamino-3-hydroxyflavone (probe F) in bilayer vesicles with the variations of fatty acid composition, polar heads, temperature, ... More
Evidence that cholesteryl ester transfer protein-mediated reductions in reconstituted high density lipoprotein size involve particle fusion.
AuthorsRye KA, Hime NJ, Barter PJ
JournalJ Biol Chem
PubMed ID9020099
'It is well established that cholesteryl ester transfer protein (CETP) changes the size of high density lipoproteins (HDL) during incubation in vitro. It has been suggested that HDL.CETP.HDL ternary complex formation is involved in these changes. The present results, which are consistent with CETP changing the size of spherical reconstituted ... More
Osmotic response in Lactobacillus casei ATCC 393: biochemical and biophysical characteristics of membrane.
AuthorsMachado MC, López CS, Heras H, Rivas EA
JournalArch Biochem Biophys
PubMed ID14725858
'The biochemical and biophysical properties of the membrane and some general characteristics of the response of Lactobacillus casei ATCC 393 (reclassified Lactobacillus zeae) to hyperosmotic conditions were studied. Under hypertonic conditions, the hydrophobicity and the bile salt sensitivity of the cultures were increased. The glycolipid AcylH3DG is only present in ... More
Agglutination of isolated rod outer segments by lectins.
AuthorsBridges CD
JournalInvest Ophthalmol Vis Sci
PubMed ID6969711
'The interactions of ten lectins with rod outer segment (ROS) plasma membranes and the effects of various haptene sugars were examined by studying agglutination in suspensions of washed ROSs. It is concluded that agglutination sites on the ROS membrane involve receptors that are recognized by eight lectins with haptene sugar ... More
Synthesis and spectral properties of a hydrophobic fluorescent probe: 6-propionyl-2-(dimethylamino)naphthalene.
AuthorsWeber G, Farris FJ
JournalBiochemistry
PubMed ID465454
'Environmentally sensitive fluorescent probes involve two groups, an electron donor and an electron acceptor, attached to an aromatic ring system, and maximal effects may be expected when these groups are as far apart as feasible. The syntheisis, characterization, and spectroscopic properties of 6-propionyl-2-(dimethylamino)naphthalene (PRODAN), a compound that fulfills these conditions, ... More
Bis-ANS as a specific inhibitor for microtubule-associated protein induced assembly of tubulin.
AuthorsMazumdar M, Parrack PK, Mukhopadhyay K, Bhattacharyya B
JournalBiochemistry
PubMed ID1633159
'5,5''-Bis[8-(phenylamino)-1-naphthalenesulfonate] (bis-ANS), the fluorescent probe which binds to tubulin, inhibits its assembly into microtubules [Horowitz et al. (1984) J. Biol. Chem. 259, 14647-14650]. The results described in this paper demonstrate that bis-ANS is quite distinct from other well-known microtubule inhibitors in its specificity of action. The inhibitory potentials of bis-ANS ... More
Fluorophores in a polar medium: time dependence of emission spectra detected by multifrequency phase and modulation fluorometry.
AuthorsParasassi T, Conti F, Gratton E
JournalCell Mol Biol
PubMed ID3948209
Inclusion complexes of ploy-beta-cyclodextrin: a model for pressure effects upon ligand-protein complexes.
AuthorsTorgerson PM, Drickamer HG, Weber G
JournalBiochemistry
PubMed ID465455
Monitoring biophysical properties of lipid membranes by environment-sensitive fluorescent probes.
AuthorsDemchenko AP, Mély Y, Duportail G, Klymchenko AS,
JournalBiophys J
PubMed ID19413953
We review the main trends in the development of fluorescence probes to obtain information about the structure, dynamics, and interactions in biomembranes. These probes are efficient for studying the microscopic analogs of viscosity, polarity, and hydration, as well as the molecular order, environment relaxation, and electrostatic potentials at the sites ... More
Interaction of Prodan with tubulin. A fluorescence spectroscopic study.
AuthorsMazumdar M, Parrack PK, Bhattacharyya B
JournalEur J Biochem
PubMed ID1740122
The compound 6-propionyl-2-(N,N-dimethyl)-aminonaphthalene (Prodan), an efficient fluorescent probe for proteins, is shown to bind to tubulin. Detailed experiments on fluorescence enhancement, anisotropy and energy transfer were carried out to unravel the nature of Prodan-tubulin interaction and the Prodan-binding site on tubulin. It was found that Prodan binds to tubulin at ... More
Binding and relaxation behaviour of prodan and patman in phospholipid vesicles: a fluorescence and 1H NMR study.
AuthorsHutterer R, Schneider FW, Sprinz H, Hof M
JournalBiophys Chem
PubMed ID8956486
The relative location, binding behaviour and the solvent relaxation behaviour of the polarity sensitive membrane probes 6-propionyl-2-(dimethylamino)naphthalene and 6-palmitoyl-2-[[trimethylammoniumethyl]methylamino]naphthalene chloride in vesicles composed of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine or egg yolk lecithin have been compared using steady-state and time-resolved fluorescence as well as high resolution NMR measurements. The reconstructed time-resolved emission spectra show ... More
Synthesis, spectral properties, and use of 6-acryloyl-2-dimethylaminonaphthalene (Acrylodan). A thiol-selective, polarity-sensitive fluorescent probe.
AuthorsPrendergast FG, Meyer M, Carlson GL, Iida S, Potter JD
JournalJ Biol Chem
PubMed ID6408077
Fluorophores processing a 6-acyl-2-dimethylaminonaphthalene moiety show fluorescence that is extremely sensitive to solvent polarity (Weber, G., and Farris, F. J. (1979) Biochemistry 18, 3075-3078). We have synthesized and characterized 6-acryloyl-2-dimethylaminonaphthalene (Acrylodan) which selectively labels thiol moieties in proteins. The quantum yield of this agent is markedly enhanced after reaction with ... More
Absorption and fluorescence of PRODAN in phospholipid bilayers: a combined quantum mechanics and classical molecular dynamics study.
AuthorsCwiklik L, Aquino AJ, Vazdar M, Jurkiewicz P, Pittner J, Hof M, Lischka H,
JournalJ Phys Chem A
PubMed ID21910413
Absorption and fluorescence spectra of PRODAN (6-propionyl-2-dimethylaminonaphthalene) were studied by means of the time-dependent density functional theory and the algebraic diagrammatic construction method. The influence of environment, a phosphatidylcholine lipid bilayer and water, was taken into account employing a combination of quantum chemical calculations with empirical force-field molecular dynamics simulations. ... More
Estimation of the polarity of the protein interior by optical spectroscopy.
AuthorsMacgregor RB, Weber G
JournalNature
PubMed ID3941741
Crystallographic studies of myoglobin have shown that the haem pocket is lined with nonpolar amino-acid residues. In order to estimate the true polarity of the interior of proteins, certain studies have used bound fluorophores, the spectroscopic properties of which reflect the polarity of their environment. These studies have most often ... More
Binding of prothrombin and its fragment 1 to phospholipid membranes studied by the solvent relaxation technique.
AuthorsHutterer R, Schneider FW, Hermens WT, Wagenvoord R, Hof M
JournalBiochim Biophys Acta
PubMed ID9804936
The phospholipid headgroup mobility of small unilamellar vesicles composed of different mixtures of phosphatidyl-L-serine (PS) and phosphatidylcholine is characterized by the solvent relaxation behavior of the polarity sensitive dyes 6-propionyl-2-(dimethylamino)naphthalene (Prodan) and 6-palmitoyl-2-[trimethylammoniumethyl]-methylamino]naphthalene chloride (Patman). If the PS content exceeds 10%, the addition of calcium leads to a substantial deceleration ... More
Two photon fluorescence microscopy of coexisting lipid domains in giant unilamellar vesicles of binary phospholipid mixtures.
AuthorsBagatolli LA, Gratton E
JournalBiophys J
PubMed ID10620293
Images of giant unilamellar vesicles (GUVs) formed by different phospholipid mixtures (1,2-dipalmitoyl-sn-glycero-3-phosphocholine/1, 2-dilauroyl-sn-glycero-3-phosphocholine (DPPC/DLPC) 1:1 (mol/mol), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine/1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPE/DPPC), 7:3 and 3:7 (mol/mol) at different temperatures were obtained by exploiting the sectioning capability of a two-photon excitation fluorescence microscope. 6-Dodecanoyl-2-dimethylamino-naphthalene (LAURDAN), 6-propionyl-2-dimethylamino-naphthalene (PRODAN), and Lissamine rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (N-Rh-DPPE) ... More
Perfusion chromatography purification of a 15 kDa rice prolamin.
AuthorsLosso JN, Bansode RR, Bawadi HA
JournalJ Agric Food Chem
PubMed ID14611182
Prolamin extracted from rice flour using 55% n-propanol contained protein impurities. Reverse phase high-performance liquid chromatography (HPLC) on a perfusion column R2/H was used to separate rice prolamin from other proteins in less than 5 min. Prolamin eluted as the major peak. The isolated prolamin migrated as a single band ... More
p36, the major cytoplasmic substrate of src tyrosine protein kinase, binds to its p11 regulatory subunit via a short amino-terminal amphiphatic helix.
AuthorsJohnsson N, Marriott G, Weber K
JournalEMBO J
PubMed ID2973411
Protein I is a hetero-tetramer which contains two copies each of p11 and p36. p36 (calpactin I, lipocortin II) is a major substrate of retrovirally encoded tyrosine protein kinases, while p11 modulates several Ca2+-induced properties also displayed by p36 alone. Here we have characterized the p11 binding site on p36 ... More
Effects of hydrostatic pressure on the location of PRODAN in lipid bilayers: a FT-IR study.
AuthorsChong PL, Capes S, Wong PT
JournalBiochemistry
PubMed ID2605189
The effects of hydrostatic pressure on the location of 6-propionyl-2-(dimethylamino)naphthalene (PRODAN), an environmentally sensitive fluorescent probe, in phosphatidylcholine lipid bilayers have been studied by Fourier-transform infrared spectroscopy (FT-IR) over the pressure range of 0.001-25 kbar. The results derived from the PRODAN C = O stretching band, the correlation field splitting ... More
Acrylodan can label amino as well as sulfhydryl groups: results with low-density lipoprotein, lipoprotein[a], and lipid-free proteins.
Human plasma lipoprotein[a] and autologous low-density lipoprotein were reacted with the fluorescent probe 6-acryloyl-2-(dimethylamino)naphthalene (acrylodan) previously reported to be specific for sulfhydryl groups. Reaction kinetics were biphasic in both cases. The reaction of bovine serum albumin with acrylodan was also biphasic. Monophasic kinetics were observed when protein free sulfhydryl groups ... More
Surface properties of cholesterol-containing membranes detected by Prodan fluorescence.
AuthorsKrasnowska EK, Bagatolli LA, Gratton E, Parasassi T
JournalBiochim Biophys Acta
PubMed ID11286976
The fluorescent membrane probe 6-propionyl-2-dimethylaminonaphthalene (Prodan) displays a high sensitivity to the polarity and packing properties of lipid membrane. Contrary to 6-lauroyl-2-dimethylaminonaphthalene (Laurdan), Prodan can also monitor the properties of the membrane surface, i.e., the polar-head pretransition. In bilayers composed of coexisting gel and liquid-crystalline phases, Prodan shows a preferential ... More
Partition of the organochlorine insecticide lindane into the human sperm surface induces membrane depolarization and Ca2+ influx.
AuthorsSilvestroni L, Fiorini R, Palleschi S
JournalBiochem J
PubMed ID9032455
The effects of the insecticide lindane (the gamma-isomer of 1,2,3,4,5,6-hexachlorocyclohexane) on membrane potential, cytosolic free Ca2+ concentration ([Ca2+]i) and surface biophysical properties were studied in human spermatozoa. The insecticide induces rapid, transient and reproducible membrane depolarization and opening of voltage-dependent Ca2+ channels leading to an increase in [Ca2+]i. In contrast ... More
Spectral properties of environmentally sensitive probes associated with horseradish peroxidase.
AuthorsLasagna M, Vargas V, Jameson DM, Brunet JE
JournalBiochemistry
PubMed ID8547280
The environmentally sensitive fluorescent probes 6-propionyl-2-(N,N-dimethylamino)naphthalene (PRODAN) and 2'-(N,N-dimethylamino)-6-naphthoyl-4-trans-cyclohexanioc acid (DANCA) form complexes with the heme binding site of apohorseradish peroxidase. The dissociation constants of the PRODAN and DANCA complexes were determined from anisotropy titration data to be approximately 8.7 x 10(-5) and 3.3 x 10(-4) M, respectively. From comparison ... More
Enhancement of Agkistrodon piscivorus piscivorus venom phospholipase A2 activity toward phosphatidylcholine vesicles by lysolecithin and palmitic acid: studies with fluorescent probes of membrane structure.
AuthorsSheffield MJ, Baker BL, Li D, Owen NL, Baker ML, Bell JD
JournalBiochemistry
PubMed ID7794890
The activity of phospholipase A2 from snake venom to hydrolyze bilayers of phosphatidylcholines is greatly enhanced by the presence of the hydrolysis products, lysolecithin and fatty acid, in the bilayer. The fluorescence of several probes of membrane structure was used to monitor changes in bilayer physical properties during vesicle hydrolysis. ... More
Ligand interactions in the adenosine nucleotide-binding domain of the Hsp90 chaperone, GRP94. II. Ligand-mediated activation of GRP94 molecular chaperone and peptide binding activity.
AuthorsWassenberg JJ, Reed RC, Nicchitta CV
JournalJ Biol Chem
PubMed ID10816560
The N-terminal domain of eukaryotic Hsp90 proteins contains a conserved adenosine nucleotide binding pocket that also serves as the binding site for the Hsp90 inhibitors geldanamycin and radicicol. Although this domain is essential for Hsp90 function, the molecular basis for adenosine nucleotide-dependent regulation of GRP94, the endoplasmic reticulum paralog of ... More
Substrate-induced translocation of PKC-alpha to the membrane.
AuthorsBruins RH, Epand RM
JournalArch Biochem Biophys
PubMed ID8554312
The protein kinase C-alpha (PKC-alpha)-catalyzed phosphorylation of the peptide [Arg]4-Tyr-Gly-Ser-[Arg]5-Tyr is independent of Ca2+ and phospholipid. The binding of this peptide to PKC-alpha induces a conformational change in the enzyme that results in the exposure of hydrophobic groups that subsequently insert into a membrane. Induction of a conformational change in ... More
Interactions of thionin from Pyrularia pubera with dipalmitoylphosphatidylglycerol large unilamellar vesicles.
AuthorsHuang W, Vernon LP, Hansen LD, Bell JD
JournalBiochemistry
PubMed ID9062115
The peptide toxin thionin from Pyrularia pubera binds to dipalmitoylphosphatidylglycerol (DPPG) large unilamellar vesicles as shown by an increase in the intensity and blue-shift of the fluorescence emission spectrum of the single tryptophan residue of the protein. The magnitude of these fluorescence changes increased with temperature near the thermotropic phase ... More
Physiological monitoring of optically trapped cells: assessing the effects of confinement by 1064-nm laser tweezers using microfluorometry.
AuthorsLiu Y, Sonek GJ, Berns MW, Tromberg BJ
JournalBiophys J
PubMed ID8889192
We report the results of microfluorometric measurements of physiological changes in optically trapped immotile Chinese hamster ovary cells (CHOs) and motile human sperm cells under continuous-wave (CW) and pulsed-mode trapping conditions at 1064 nm. The fluorescence spectra derived from the exogenous fluorescent probes laurdan, acridine orange, propidium iodide, and Snarf ... More
Properties of palmitoyl phosphatidylcholine, sphingomyelin, and dihydrosphingomyelin bilayer membranes as reported by different fluorescent reporter molecules.
AuthorsNyholm T, Nylund M, Söderholm A, Slotte JP
JournalBiophys J
PubMed ID12547780
The properties of vesicle membranes prepared from 16:0-SM, 16:0-DHSM, or DPPC were characterized using steady-state and time-resolved fluorescence spectroscopy and different fluorescent reporter molecules. The acyl-chain region was probed using free and phospholipid-bound 1,6-diphenyl-1,3,5-hexatriene. 16:0-DHSM was found to be the more ordered than both DPPC and 16:0-SM 5 degrees C ... More
Cooperative transition in the conformation of 24-mer tarantula hemocyanin upon oxygen binding.
AuthorsErker W, Beister U, Decker H
JournalJ Biol Chem
PubMed ID15695808
Hemocyanins are large respiratory proteins of arthropods and mollusks, which bind oxygen with very high cooperativity. Here, we investigated the relationship between oxygen binding and structural changes of the 24-mer tarantula hemocyanin. Oxygen binding of the hemocyanin was detected following the fluorescence intensity of the intrinsic tryptophans. Under the same ... More
The influence of apolipoproteins on the structure and function of spheroidal, reconstituted high density lipoproteins.
AuthorsRye KA, Barter PJ
JournalJ Biol Chem
PubMed ID8144610
This study investigates the influence of apolipoproteins on the structure and function of spheroidal, reconstituted high density lipoproteins (rHDL). Spheroidal rHDL containing apolipoprotein (apo) A-I, (A-I)rHDL, were prepared by incubating discoidal rHDL with low density lipoproteins and lecithin:cholesterol acyltransferase. Spheroidal rHDL containing apoA-II, (A-II)rHDL, were prepared by displacing apoA-I from ... More
The mechanism of the remodeling of high density lipoproteins by phospholipid transfer protein.
AuthorsSettasatian N, Duong M, Curtiss LK, Ehnholm C, Jauhiainen M, Huuskonen J, Rye KA
JournalJ Biol Chem
PubMed ID11325961
Phospholipid transfer protein (PLTP) remodels high density lipoproteins (HDL) into large and small particles. It also mediates the dissociation of lipid-poor or lipid-free apolipoprotein A-I (apoA-I) from HDL. Remodeling is enhanced markedly in triglyceride (TG)-enriched HDL (Rye, K.-A., Jauhiainen, M., Barter, P. J., and Ehnholm. C. (1998) J. Lipid. Res. ... More
Preferential interactions of fluorescent probe Prodan with cholesterol.
AuthorsBondar OP, Rowe ES
JournalBiophys J
PubMed ID9916026
The fluorescent probe Prodan has been widely used as a probe of model and biological membranes. Its fluorescent maxima in phospholipid bilayers vary as a function of phase state, with maxima at 485 for the liquid crystal Lalpha, 435 nm for the gel L'beta, and 507 nm for the interdigitated ... More
Calmodulin-dependent enzymes undergo a protein-induced conformational change that is associated with their interactions with calmodulin.
AuthorsHuang S, Carlson GM, Cheung WY
JournalJ Biol Chem
PubMed ID8125988
The anionic hydrophobic (amphipathic) fluorescent probe 2-(p-toluidinyl)-naphthalene-6-sulfonate was used to investigate the surface hydrophobic properties of calmodulin (CaM)-dependent enzymes as follows: calcineurin, myosin light chain kinase, cyclic nucleotide phosphodiesterase, CaM-dependent protein kinase II, and the gamma-subunit of phosphorylase kinase. We found that certain domains of these enzymes that interacted with ... More
A two-photon view of an enzyme at work: Crotalus atrox venom PLA2 interaction with single-lipid and mixed-lipid giant unilamellar vesicles.
AuthorsSanchez SA, Bagatolli LA, Gratton E, Hazlett TL
JournalBiophys J
PubMed ID11916878
We describe the interaction of Crotalus atrox-secreted phospholipase A2 (sPLA2) with giant unilamellar vesicles (GUVs) composed of single and binary phospholipid mixtures visualized through two-photon excitation fluorescent microscopy. The GUV lipid compositions that we examined included 1-palmitoyl-2-oleoyl-phosphatidylcholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) (above their gel-liquid crystal transition temperatures) and two ... More
Differential-wavelength deconvolution of time-resolved fluorescence intensities. A new method for the analysis of excited-state processes.
AuthorsLakowicz JR, Balter A
JournalBiophys Chem
PubMed ID7171715
We describe a new procedure for the analysis of time-resolved decays of fluorescence intensity observed for excited-state reactions. This procedure is particularly valuable because it simplifies both determination of the rate constants of excited-state reactions and calculation of the emission spectra of the unreacted and the reacted species. These advantages ... More
Sphingomyelin inhibits the lecithin-cholesterol acyltransferase reaction with reconstituted high density lipoproteins by decreasing enzyme binding.
AuthorsBolin DJ, Jonas A
JournalJ Biol Chem
PubMed ID8702592
Lecithin-cholesterol acyltransferase (LCAT) catalyzes the formation of cholesterol esters on high density lipoproteins (HDL) and plays a critical role in reverse cholesterol transport. Sphingomyelin, an important constituent of HDL, may regulate the activity of LCAT at any of the key steps of the enzymatic reaction: binding of LCAT to the ... More
Effects of alcohol-induced lipid interdigitation on proton permeability in L-alpha-dipalmitoylphosphatidylcholine vesicles.
AuthorsZeng J, Smith KE, Chong PL
JournalBiophys J
PubMed ID8274634
6-Carboxyfluorescein was employed to examine the effect of alcohol-induced lipid interdigitation on proton permeability in L-alpha-dipalmitoylphosphatidylcholine (DPPC) large unilamellar vesicles. Proton permeability was measured by monitoring the decrease of 6-carboxyfluorescein fluorescence after a pH gradient from 3.5 (outside the vesicle) to 8.0 (inside the vesicle) was established. At 20 degrees ... More
Interactions between pressure and ethanol on the formation of interdigitated DPPC liposomes: a study with Prodan fluorescence.
AuthorsZeng JW, Chong PL
JournalBiochemistry
PubMed ID1892848
Steady-state fluorescence of 6-propionyl-2-(dimethylamino)naphthalene (Prodan) has been employed to study the interacting effects between ethanol and pressure on the formation of the fully interdigitated dipalmitoylphosphatidylcholine (DPPC). At 1 atm and 20 degrees C, a dramatic change in the emission spectrum of Prodan fluorescence is observed at about 1.1-1.3 M ethanol. ... More
Surface and micellar properties of new nonionic gemini aldonamide-type surfactants.
AuthorsKomorek U, Wilk KA
JournalJ Colloid Interface Sci
PubMed ID14757096
A new group of gemini aldonamide-type surfactants-N,N'-bisalkyl-N,N'-bis[(3-gluconylamide)propyl]ethylenediamines, N,N'-bisdodecyl-N,N'-bis[(3-glucoheptonylamide)propyl]ethylenediamine, and N,N'-bisalkyl-N,N'-bis[(3-lactobionylamide)propyl]ethylenediamines, (alkyl: n-C(8)H(17), n-C(12)H(25)), were synthesized and characterized. The surface properties, such as surface excess concentration, Gamma(cmc), surface area demand per molecule, A(min), efficiency in surface tension reduction, pC(20), the effectiveness of surface tension reduction, gamma(cmc), critical micelle concentration, cmc, and ... More
Effects of hydrostatic pressure on the location of PRODAN in lipid bilayers and cellular membranes.
AuthorsChong PL
JournalBiochemistry
PubMed ID3349041
The effects of hydrostatic pressure on the location of 6-propionyl-2-(dimethylamino)naphthalene (PRODAN), an environmentally sensitive fluorescent probe, in phosphatidylcholine lipid bilayers and in goldfish brain synaptic membranes have been studied by fluorescence spectroscopy over the pressure range of 0.001-2 kbar. The emission spectrum of PRODAN in all the membrane systems examined ... More
Interfacial properties of model membranes and plasma lipoproteins containing ether lipids.
AuthorsMassey JB, She HS, Pownall HJ
JournalBiochemistry
PubMed ID4074734
The interfacial properties of synthetic ester and ether phosphatidylcholines (PCs) were investigated by using the polarity-sensitive fluorescent probes 6-propionyl-2-(dimethylamino)naphthalene (Prodan) and pyrene. The physical state of the phospholipid matrix was determined by fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH). Single-bilayer phospholipid vesicles formed by sonication and model high-density lipoproteins were studied. On ... More
Effect of ethanol-induced lipid interdigitation on the membrane solubility of Prodan, Acdan, and Laurdan.
AuthorsZeng J, Chong PL
JournalBiophys J
PubMed ID7696509
The effect of ethanol-induced lipid interdigitation on the partition coefficient (Kp) of 6-propionyl-2-(dimethylamino)naphthalene (Prodan) and its two derivatives, 6-acetyl-2-(dimethylamino)naphthalene (Acdan) and 6-lauroyl-2-(dimethylamino)naphthalene (Laurdan), in L-alpha-dipalmitoylphosphatidylcholine (DPPC) vesicles has been examined by a precipitation method over the ethanol concentration range of 0-1.8 M. At 20 degrees C and in the absence ... More
Polarity decrease at the adhesive junction between two model membranes containing gangliosides.
AuthorsBrewer GJ
JournalBiochemistry
PubMed ID1737033
The increased electrical conductance previously observed between two model membranes containing gangliosides suggests the creation of a new environment in the adhesive junction [Brewer, G. J., & Thomas, P.D. (1984) Biochim. Biophys. Acta 776, 279]. In order to provide a mechanism for this novel finding, we now report an investigation ... More
Caged protein conjugates and light-directed generation of protein activity: preparation, photoactivation, and spectroscopic characterization of caged G-actin conjugates.
AuthorsMarriott G
JournalBiochemistry
PubMed ID8049211
A simple method is described to prepare caged (inactive) protein complexes using the amino group-directed photo-deprotection group [(nitroveratryl)oxy]chlorocarbamate (NVOC-Cl). In this study, I show how the polymerization activity of G-actin in physiological salt solution is lost upon conjugation of essential lysine residues of G-actin with NVOC-Cl. Reaction conditions were optimized ... More
A search for site-filling ligands in the Mcg Bence-Jones dimer: crystal binding studies of fluorescent compounds.
AuthorsEdmundson AB, Ely KR, Herron JN
JournalMol Immunol
PubMed ID6431268
In trigonal crystals grown in 1.9 M ammonium sulfate buffered at pH 6.2, the Mcg light-chain (Bence-Jones) dimer has a highly aromatic binding cavity accessible to a wide range of hydrophobic and aromatic ligands. A search was made for site-filling ligands by diffusing compounds into the crystals and determining their ... More
Prodan fluorescence reflects differences in nucleotide-induced conformational states in the myosin head and allows continuous visualization of the ATPase reactions.
AuthorsHiratsuka T
JournalBiochemistry
PubMed ID9585528
The noncovalent fluorescent probe 6-propionyl-2-(dimethylamino)naphthalene (prodan) binds stoichiometrically to myosin subfragment-1 (S-1) without affecting the ATPase and actin-binding properties of S-1. Neither ATP nor actin interferes with the prodan binding. Free prodan exhibits a green emission peak at 520 nm. However, the prodan bound to S-1 and the S-1.ADP complex ... More
Spatial relationship between the prodan site, Trp-214, and Cys-34 residues in human serum albumin and loss of structure through incremental unfolding.
AuthorsKrishnakumar SS, Panda D
JournalBiochemistry
PubMed ID12044178
Prodan (6-propionyl-2-(dimethylamino)-naphthalene), a competitive inhibitor of warfarin binding to human serum albumin (HSA) at drug site I, was used to determine the inter- and intradomain distances of HSA. The fluorescence resonance energy transfer (FRET) distances between prodan and Trp-214, prodan and 7-(diethyl amino)-4-methylcoumarin 3-maleimide (CM)-modified Cys-34, and Trp-214 and CM-Cys-34 ... More