Measurement of transmembrane potentials in phospholipid vesicles.
AuthorsKornberg RD, McNamee MG, McConnell HM
JournalProc Natl Acad Sci U S A
PubMed ID4338594
Phosphatidylcholine vesicles are permeable to tempotartrate, a spin-label derivative of tartaric acid. The inside-outside distribution of tempotartrate is coupled to the inside-outside distribution of H(+), so it must be a measure of the transmembrane electrical potential difference in vesicles permeable to H(+). This prediction is borne out by the finding ... More
Calcium translocation mechanism in sarcoplasmic reticulum vesicles, deduced from location studies of protein-bound spin labels.
AuthorsChampeil P, Rigaud JL, Gary-Bobo CM
JournalProc Natl Acad Sci U S A
PubMed ID6446710
'Sarcoplasmic reticulum vesicles were exposed to various thiol-directed spin labels, and the position of the label on the inner or outer vesicle surface was investigated as a function of the ATPase (adenosinetriphosphatase; ATP phosphohydrolase, EC 3.6.1.3) chemical state. Previous measurements of label accessibility to externally added ascorbate had been considered ... More
Location of diphenylhexatriene (DPH) and its derivatives within membranes: comparison of different fluorescence quenching analyses of membrane depth.
AuthorsKaiser RD, London E
JournalBiochemistry
PubMed ID9609714
'The average membrane location of a series of diphenylhexatriene (DPH)-derived membrane probes was analyzed by measuring the quenching of DPH fluorescence with a series of nitroxide-labeled lipids in which the depth of the nitroxide group is varied. All DPH derivatives were located deeply within the bilayer. Some derivatives were anchored ... 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
Spin label partitioning in lipid vesicles. A model study for drug encapsulation.
AuthorsDefrise-Quertain F, Chatelain P, Ruysschaert JM, Delmelle M
JournalBiochim Biophys Acta
PubMed ID6243999
'In this paper we attempt to outline some features which determine the encapsulation of small molecules into lipid vesicles. Spin labels derived from five carboxylic acids of different lengths were synthesized and incorporated in varying amounts into multilamellar and unilamellar vesicles made up of four different phosphatidylcholines. The influence on ... More
Effect of fatty acyl chain length of phosphatidylcholine on their transfer from liposomes to erythrocytes and transverse diffusion in the membranes inferred by TEMPO-phosphatidylcholine spin probes.
AuthorsTamura A, Yoshikawa K, Fujii T, Ohki K, Nozawa Y, Sumida Y
JournalBiochim Biophys Acta
PubMed ID3004580
'TEMPO-phosphatidylcholine (PC) spin probes which have homologous saturated acyl chains of 10, 12, 14 and 16 carbon atoms, were synthesized as analogues of PC. Transfer of TEMPO-PCs from liposomal membrane to the ghost membrane of human erythrocyte and transverse diffusion of TEMPO-PCs within the membrane of intact erythrocytes were determined ... More
Control of the depth of molecules within membranes by polar groups: determination of the location of anthracene-labeled probes in model membranes by parallax analysis of nitroxide-labeled phospholipid induced fluorescence quenching.
AuthorsAsuncion-Punzalan E, London E
JournalBiochemistry
PubMed ID7547874
'The location of anthracene-labeled molecules incorporated into model membranes was measured by fluorescence quenching. The depth of the anthracene group was calculated from the degree of quenching by lipids carrying a nitroxide at different depths, using the parallax analysis (Chattopadhyay & London (1987) Biochemistry 26, 39-45). A series of anthracene ... More
A new method of evaluating complement mediated lysis of liposomes.
AuthorsVistnes AI
JournalJ Immunol Methods
PubMed ID6323583
'Immune lysis of liposomes has been utilized in immunoassays for detection of specific antibodies. Membrane immunoassays which have sensitivities comparable with those of radioimmunoassays are easy and rapid to perform. Information obtained so far has been limited by the fact that the method is based on empirical relationships. The present ... More
Oxidation of hydroxylamines to nitroxide spin labels in living cells.
AuthorsChen K, Swartz HM
JournalBiochim Biophys Acta
PubMed ID2840969
'In the presence of oxygen, cells can oxidize hydroxylamines, which are the products of the reduction of nitroxides in cells, back to nitroxides. Lipid-soluble hydroxylamines are oxidized much more rapidly than water-soluble ones, and most of this oxidation is inactivated by heat or trichloroacetic acid, indicating that the principal mechanism ... More
Dynamic fluorescence quenching studies on lipid mobilities in phosphatidylcholine-cholesterol membranes.
AuthorsMerkle H, Subczynski WK, Kusumi A
JournalBiochim Biophys Acta
PubMed ID3028480
'Bimolecular collision rate of 8-anilinonaphthalene-1-sulfonic acid (ANS) and the nitroxide doxyl group attached to various carbons on stearic acid spin labels (n-SASL) in phosphatidylcholine-cholesterol membranes in the fluid phase was studied by observing dynamic quenching of ANS fluorescence by n-SASL's. The excited-state lifetime of ANS and its reduction by the ... More
Reactive oxygen species play no role in the candidacidal activity of the salivary antimicrobial peptide histatin 5.
AuthorsVeerman EC, Nazmi K, Van't Hof W, Bolscher JG, Den Hertog AL, Nieuw Amerongen AV
JournalBiochem J
PubMed ID15109304
'The mechanism of action of antimicrobial peptides is still a matter of debate. The formation of ROS (reactive oxygen species) has been suggested to be the crucial step in the fungicidal mechanism of a number of antimicrobial peptides, including histatin 5 and lactoferrin-derived peptides. In the present study we have ... More
Spin labeled acetylcholine analogs: studies of cholinergic receptor.
AuthorsRosen GM, Abou-Donia MB, Yeh JZ, Menzel DB
JournalRes Commun Chem Pathol Pharmacol
PubMed ID172996
'Some spin-labeled acetylcholine analogs, in which the number of methylene groups between the quaternary nitrogen and the ether oxygen ranged between 1-5, were synthesized to study drug interacitons with acetylcholine receptors. None of the compounds tested, with the exception of the one that contained 2 methylene groups (SL-2) had any ... More
Effect of hyperthermia on lymphocyte plasma membrane. 3 electrolyte spin labels permeability.
AuthorsKoter M
JournalCytobios
PubMed ID2591253
'The effect of elevated temperatures on the transport of electrolyte spin labels anionic c-TEMPIR and cationic TEMPO-choline character across the porcine lymphocyte plasma membrane was investigated. Breaks in the Arrhenius plot for permeability of both spin-labels occurred at 42 to 43 degrees C. TEMPO-choline and c-TEMPIR transport are probably critical ... More
Characterization of dimyristoylphosphatidylcholine vesicles and their dimensional changes through the phase transition: molecular control of membrane morphology.
AuthorsWatts A, Marsh D, Knowles PF
JournalBiochemistry
PubMed ID656398
Spin membrane immunoassay for use in meningococcal serology.
AuthorsVistnes AI, Rosenqvist E, Frøholm LO
JournalJ Clin Microbiol
PubMed ID6313752
'A modified and improved spin membrane immunoassay has been developed for detecting complement-activating antibodies to Neisseria meningitidis capsular polysaccharide antigens. The polysaccharides were incorporated in the membranes of large unilamellar vesicles prepared by the reverse-phase evaporation method and filled with the water-soluble spin label tempocholine chloride. Upon addition of group-specific ... More
Physical and chemical interactions between nitric oxide and nitroxides.
AuthorsSingh RJ, Hogg N, Mchaourab HS, Kalyanaraman B
JournalBiochim Biophys Acta
PubMed ID7803474
'The physical and chemical interaction of nitric oxide (NO) with stable nitroxides have been studied in both aqueous and membrane environments. The ESR spectrum of 3-carbamoyl-2,2,5,5-tetramethyl-3-pyrroline-1- yloxy (CTPO) was observed to broaden upon exposure to NO. This effect can be explained by invoking Heisenberg spin exchange as has been previously ... More
Effect of gamma radiation on the transport of spin-labeled compounds across the erythrocyte membrane.
AuthorsGwozdzinski K, Bartosz G, Leyko W
JournalRadiat Environ Biophys
PubMed ID7280184
'The effect of ionizing radiation on the non-electrolyte, anion and cation permeability of the erythrocyte membrane was studied by measurement of the reduction rate of appropriate nitroxyl derivatives. Irradiation of bovine erythrocytes in the dose-range of 2-50 krad resulted in a regular dose-dependent increase in the reduction rates of a ... More
Evidence for phase boundary lipid. Permeability of Tempo-choline into dimyristoylphosphatidylcholine vesicles at the phase transition.
AuthorsMarsh D, Watts A, Knowles PF
JournalBiochemistry
PubMed ID182212
'The existence of distinct regions of mismatch in molecular packing at the interfaces of the fluid and ordered domains during the phase transition of dimyristoylphosphatidylcholine vesicles has been demonstrated by measuring the temperature dependence of the permeability to a spin-label cation and comparing this with a statistical mechanical calculation of ... More
Permeation of a spin-label phosphate into the human erythrocyte.
AuthorsRoss AH, McConnell HM
JournalBiochemistry
PubMed ID168918
'The reduction of spin-labels by human erythrocytes can be used to follow their penetration into these cells. The neutral spin-label alcohol Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidinyl-1-oxyl) diffuses through the membrane very quickly. The membrane is virtually impermeable to the positively charged spin-label Tempo-choline (N,N-dimethyl-N-(2'',2'',6'',6''-tetramethyl-4''-piperidinyl-1-oxyl)-2-hydroxyethylammonium chloride). The negatively charged spin-label Tempo phosphate (4-phospho-2,2,6,6-tetramethylpiperidinyl-l-oxyl) is ... More
Asymmetric permeability of the membrane of egg phosphatidylcholine-cholesterol vesicles to 2,2,6,6 tetramethyl piperidinyl-l-oxycholine.
AuthorsSetaka M, Ichiki T, Shimizu H
JournalJ Biochem (Tokyo)
PubMed ID207683
'The rates of uptake and release of 2,2,6,6 tetramethyl piperidinyl-l-oxycholine (Tempo-choline) for vesicles made of dipalmitoyl phosphatidylcholine (DPPC) and of egg phosphatidylcholine-cholesterol mixtures were measured by ESR and found to have interesting temperature-dependences. In the former case, both rates exhibit a sharp maximum at the critical temperature of phase transition ... More
Membrane immunoassay: a spin membrane immunoassay for thyroxine.
AuthorsTan CT, Chan SW, Hsia JC
JournalMethods Enzymol
PubMed ID6275239
Inside-outside transitions of phospholipids in vesicle membranes.
AuthorsKornberg RD, McConnell HM
JournalBiochemistry
PubMed ID4324203
Synthesis of some spin-labeled analogs of drug molecules.
AuthorsChignell CF, Starkweather DK, Erlich RH
JournalJ Med Chem
PubMed ID4339773
Transmembrane potentials and phospholipid flip-flop in excitable membrane vesicles.
AuthorsMcNamee MG, McConnell HM
JournalBiochemistry
PubMed ID4354249
Infection-triggered release of tempocholine from bacteriophage G4 studied by electron spin resonance.
AuthorsMaeda T, Ohnishi SI, Yanagida M
JournalFEBS Lett
PubMed ID207568
Effects of Ca2+ and membrane surface charge on the direct lytic activity of cobra cardiotoxin - a membrane spin assay.
AuthorsHsia JC, Er SS, Lee CY
JournalBiochem Biophys Res Commun
PubMed ID203288
A ganglioside spin label: ganglioside head group interactions.
AuthorsSharom FJ, Grant CW
JournalBiochem Biophys Res Commun
PubMed ID191002
Binding of antibodies to nitroxide spin labels and to the corresponding hydroxylamines.
AuthorsRey P, McConnell HM
JournalBiochem Biophys Res Commun
PubMed ID1033765
Stearoyl paratoluenesulfonate. A powerful acylating agent for lipid synthesis.
AuthorsKeana JF, Ertle AR
JournalChem Phys Lipids
PubMed ID991389
The utility of the mixed carboxylic-sulfonic acid anhydride stearoyl p-toluenesulfonate as a powerful, mild acylating agent for lipid synthesis is shown by the synthesis of rac 1,2-distearoyl-3-iodopropane, lecithin and a spin-labeled choline derivative from the corresponding alcohols. The method constitutes a significant improvement of earlier acylating methods. ... More
Immune reactivity against membranes containing ganglioside GM1 in chronic-progressive multiple sclerosis: observation by spin-membrane immunoassay.
AuthorsFeix JB, Khatri B, McQuillen MP, Koethe SM
JournalImmunol Commun
PubMed ID6097532
A spin-membrane immunoassay has been employed to examine the immune reactivity of whole serum from patients with chronic-progressive multiple sclerosis (CPMS) against liposomes containing ganglioside GM1. Exposure to serum resulted in complement-mediated lysis of the GM1-liposomes. No lysis occurred with liposomes devoid of ganglioside. The mean (+/- S.E.M.) lysis values ... More
Immune lysis of spin label loaded liposomes incorporating cardiolipin; a new sensitive method for detecting anticardiolipin antibodies in syphilis serology.
AuthorsRosenqvist E, Vistnes AI
JournalJ Immunol Methods
PubMed ID321698
Liposomes prepared from a mixture of the pure lipids cholesterol, lecithin and cardiolipin (molar ratio 50/45/5), are able to bind antibodies directed against Treponema pallidum. When the liposomes are loaded with the water soluble spin label tempocholine chloride, the release of spin label from the liposomes can be monitored directly ... More
Influence of structure on the reduction of nitroxide MRI contrast-enhancing agents by ascorbate.
AuthorsKeana JF, Van Nice FL
JournalPhysiol Chem Phys Med NMR
PubMed ID6537509
With an eye toward the design of new nitroxide-based magnetic resonance imaging (MRI) contrast agents, the effect of ring size and the presence of proximate polar groups on the rate of reduction of cyclic nitroxides by the biologically relevant reducing agent ascorbate ion have been determined. To the extent that ... More
Amine and carboxylate spin probe permeability in red cells.
AuthorsTodd AP, Mehlhorn RJ, Macey RI
JournalJ Membr Biol
PubMed ID2549250
Permeabilities for a homologous series of amine and carboxylate nitroxide spin probes were measured in human red blood cells by an electron paramagnetic resonance (EPR) method. Permeabilities determined in this study are much lower than would be predicted for a sheet of bulk hydrocarbon and the polarity of the rate-limiting ... More
Effects of amphotericin B on membrane permeability--kinetics of spin probe reduction.
AuthorsAracava Y, Schreier S, Phadke R, Deslauriers R, Smith IC
JournalBiophys Chem
PubMed ID6279197
The effect of the polyene antibiotic amphotericin B on the permeability of both unilamellar and multilamellar model membranes is investigated. The method measures the loss of the electron paramagnetic resonance signal of a spin probe, trapped in the aqueous compartment of a lipid dispersion, upon addition of ascorbate ions to ... More
Spin membrane immunoassay: simplicity and specificity.
AuthorsChan SW, Tan CT, Hsia JC
JournalJ Immunol Methods
PubMed ID207779
The many disadvantages of radioimmunoassay (RIA) have stimulated attempts to develop non-radioactive assays. These include spin immunoassay (SIA), which is simple, specific, and requires no separation procedures but is much less sensitive than RIA. The membrane immunoassay (MIA) described here is more sensitive than the SIA. Serum is prepared as ... More
Transport of spin-labeled tetracycline across model and biological membranes.
AuthorsPezeshk A, Pezeshk V, Firlej A, Wojas J, Subczynski WK
JournalLife Sci
PubMed ID8383269
Electron spin resonance spectra of spin labeled tetracycline (TC-SL) do not show any recognizable partitioning into a lipid bilayer or bulk hydrocarbon solvent, paraffin oil. TC-SL, however, penetrates through the model and biological membranes. It is shown that the rate of permeation depends on membrane composition and increases with temperature. ... More
Electron spin resonance spin label studies of plasma fibronectin: effect of temperature.
AuthorsLai CS, Tooney NM
JournalArch Biochem Biophys
PubMed ID6320741
Plasma fibronectin was chemically modified by 4-maleimido-2,2,6,6-tetramethylpiperidinooxyl (maleimide spin label). Only the free sulfhydryl groups of plasma fibronectin were modified by the label under the experimental conditions. The ESR spectrum of spin-labeled fibronectin showed that the sites of labeling were highly immobilized, suggesting that the sulfhydryl groups of the protein ... More
A new electron spin resonance assay for membrane asymmetry and entrapped volume of unilamellar lipid vesicles based on photoreduced flavin adenine dinucleotide.
AuthorsThurnhofer H, Kräutler B, Hauser H
JournalBiochemistry
PubMed ID2541782
A new ESR assay has been developed for the characterization of unilamellar lipid vesicles. It is based on the reduction by photogenerated FADH2 of amphiphilic spin-labels having the spin in the polar group. FADH2 is generated in situ under anaerobic conditions from its oxidized form (FAD) by photoreduction in the ... More
The efflux of spin label entrapped in human erythrocyte ghosts when suspended in hyposmolar solutions. The effect of chlorpromazine, trifluoperazine, nicardipine and some other membrane active substances.
AuthorsLagercrantz C, Larsson T, Sörskog L, Lincoln P
JournalBiochem Pharmacol
PubMed ID6329223
Human erythrocyte ghosts were loaded with the spin label tempocholine . Once entrapped in the ghosts, this spin label, carrying a positive charge, is not able to penetrate through intact ghost membranes. The ghosts were loaded with spin label to a concentration high enough to introduce exchange broadening of the ... More
Calcium ion independent membrane leakage induced by phospholipase-like myotoxins.
The two snake venom myotoxins ammodytin L and myotoxin II, purified respectively from Vipera ammodytes ammodytes and Bothrops asper, have phospholipase-like structures but lack an Asp-49 in the active site and are without normal phospholipase activity. The interaction of these proteins with different types of liposomes indicated that the myotoxins ... More
Luminescence quenching by nitroxide spin labels in aqueous solution: studies on the mechanism of quenching.
AuthorsMatko J, Ohki K, Edidin M
JournalBiochemistry
PubMed ID1731926
The mechanism of luminescence quenching by spin labels was investigated in aqueous solution by steady-state and time-resolved luminescence techniques. Water-soluble nitroxide radicals strongly quenched the luminescence emitted by Tb3+ chelates and by fluorescein, either free or conjugated to proteins. The following features of the quenching reaction were established: (I) the ... More
Extension of the parallax analysis of membrane penetration depth to the polar region of model membranes: use of fluorescence quenching by a spin-label attached to the phospholipid polar headgroup.
AuthorsAbrams FS, London E
JournalBiochemistry
PubMed ID8399232
The parallax method is a method by which the depth of fluorescent molecules within a membrane is calculated from the ratio of quenching induced by two spin-labeled phospholipids at different depths. In this report, the method is extended to measurements of depth in the polar headgroup region of the membrane ... More
Collisions between nitrogen-14 and nitrogen-15 spin-labels. 1. Lipid-lipid interactions in model membranes.
AuthorsDavoust J, Seigneuret M, Hervé P, Devaux PF
JournalBiochemistry
PubMed ID6309216
A new method is described to evaluate collision frequencies between spin-labeled molecules in membranes. This method utilizes a mixture of 14N and 15N nitroxides. The extent of spin-spin interaction between the two different isotopes can be determined by comparing experimental ESR spectra with simulated spectra. The latter are calculated by ... More
Entrapping of the spin label tempocholine into human erythrocytes by resealing after hyposmolar stress. Comparison with haemolysis. The effects of some membrane-active substances.
AuthorsLagercrantz C, Larsson T, Tollsten L, Prus K
JournalBiochem Pharmacol
PubMed ID2981529
Human erythrocytes were subjected to a sudden hyposmolar stress by suspension in solutions of varying salt concentrations in the presence of the spin label tempocholine. The enlarged pores in the erythrocyte membranes produced by the influx of water, followed by stretching, allowed the passage of the spin label, so that ... More
On the origin of sphingolipid/cholesterol-rich detergent-insoluble cell membranes: physiological concentrations of cholesterol and sphingolipid induce formation of a detergent-insoluble, liquid-ordered lipid phase in model membranes.
AuthorsAhmed SN, Brown DA, London E
JournalBiochemistry
PubMed ID9283086
Detergent-insoluble membrane fragments that are rich in sphingolipid and cholesterol can be isolated from both cell lysates and model membranes. We have proposed that these arise from membranes that are in the liquid-ordered phase both in vivo and in vitro [Schroeder et al. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, ... More
The eosin-5-maleimide binding site on human erythrocyte band 3: investigation of membrane sidedness and location of charged residues by triplet state quenching.
AuthorsPan RJ, Cherry RJ
JournalBiochemistry
PubMed ID9665731
The triplet probe eosin-5-maleimide (EMA) is a specific inhibitor of anion transport mediated by the erythrocyte membrane protein, band 3. It was previously shown that the eosin moiety is located close to the anion binding site when EMA is covalently bound to band 3 [Pan, R.-j., and Cherry, R. J. ... More
Spin label oximetry to assess extracellular oxygen during myocardial ischemia.
AuthorsBaker JE, Froncisz W, Joseph J, Kalyanaraman B
JournalFree Radic Biol Med
PubMed ID8958135
We describe real-time measurement of myocardial oxygen consumption during ischemia in the intact heart. Measurement of extracellular oxygen concentration during myocardial ischemia by spin label oximetry has been limited by ischemia-induced reduction of the neutral, water-soluble nitroxide TEMPONE. We have overcome this problem by encapsulating the nitroxides. Isolated immature (7-10 ... More