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M A Singer

Publications and source records attributed to M A Singer.

At least 55 records · Page 3Linked to original sources
PubMed1986

The metastability of saturated phosphatidylcholines depends on the acyl chain length.

We report the peak temperatures and enthalpies of the main (gel to liquid crystal) transitions and 's-transitions' of symmetric saturated PC's (CnPC) with n varying from 12 to 22, where s-transition is defined as the transition between metastable and stable phases. The line for s-transition peak temperature versus n (t = 1.98n -16.0 degrees C) crosses the corresponding pre- and main-transition curves between n = 12 and 14. We also find that the metastability pattern for lipids of n less than 14 is different to that for n greater than 14 (and is similar to that of the phosphatidylethanolamine analogs). For n greater than 14 the enthalpy of the s-transition decreases with increasing n and extrapolates to zero above n = 20, implying that the s-(orthorhombic) and gel (quasi-hexagonal) phases become identical. That simultaneous gel-liquid crystal transition and metastability is restricted to the range n = 12 to 20 may be correlated with the predominance in natural lipids of these acyl chain lengths.

Liposomes↗
PubMed1986

Effect of saturated phosphatidylcholines on the functional properties of reconstituted cytochrome oxidase.

Cytochrome oxidase was incorporated into liposomes, at various protein/lipid ratios, composed of either a phosphatidylcholine of varying chain length and symmetry or asolectin. Catalytic activity and respiratory control were assayed at two temperatures. All preparations showed higher activity at low protein/lipid ratios, but only asolectin showed respiratory control. A spectroscopic determination of the vectorial orientation of oxidase molecules showed that, for proteoliposomes with saturated lipids, 100% of oxidase molecules could be reduced by external substrate as compared with 75% for asolectin proteoliposomes. Freeze-fracture electron microscopy confirmed that oxidase was incorporated into these proteoliposomes and differential scanning calorimetry indicated that the protein induces significant disruption in the long range packing of the saturated phospholipids. We propose that the oxidase molecules in proteoliposomes formed from saturated phosphatidylcholines do not display respiratory control because they are unable to assume the transmembrane orientation necessary for full vectorial activity.

Animals↗
PubMed1985

Permeability and morphology of low temperature phases in bilayers of single and of mixtures of phosphatidylcholines.

The properties of subtransitions were studied in aqueous dispersions of saturated phosphatidylcholines (PC) by means of permeability measurements, freeze-fracture electron microscopy, and differential scanning calorimetry (DSC). For dispersions of C16PC, a C16PC analog (2,3-dipalmitoyl-cyclopentano-1-phosphocholine with four methylene residues between the nitrogen and the phosphorus atoms) and C17PC, there was good agreement between phase properties (including subtransitions) as observed by DSC and temperature-related permeability. C16PC and C17PC dispersions also displayed a 'crinkled' surface morphology in the subgel state. The phase diagram for mixtures of C14PC and C16PC was consistent with ideal mixing of these two components in the subgel state and also illustrated the relative independence of the subtransition on acyl chain length as compared to the pre- and main transitions. Together, these results indicate that (i) permeability, DSC and freeze-fracture electron microscopy measurements do correlate reasonably well with the existence of a subgel state, (ii) mixtures of lipids with similar acyl chain lengths can be used to investigate subtransitions, (iii) the development of a subtransition appears to be mainly a function of the non-acyl chain moiety of the phospholipid.

Calorimetry, Differential Scanning↗
PubMed1984

Lateral interaction of cholesterol in diacylphosphatidylcholesterol bilayers.

Thermotropic phase-transition properties of the aqueous dispersions of several diacylphosphatidylcholesterol (DRCh) analogs are examined. The aqueous dispersions of their calcium salts exhibit characteristic endothermic thermotropic transitions due to a change in the conformation of acyl chains. These dispersions consist of osmotically intact liposomes that trap ions, and at the transition temperature there is anomalous increase in the ion leakage. Wide-angle electron diffraction studies of DPCh . Ca monolayers also exhibit a transition from a sharp 4.25 A band to a broad one centering at 4.7 A, reflecting an order-disorder transition in the acyl chains. The long-range order in the organization of acyl chains of DRCh molecules could arise from intermolecular interactions between the cholesterol moieties to form a functional dimer, and such dimers are apparently cross-linked by Ca2+ to form a long-range interacting lattice of acyl chains. Evidence for this model is adduced from the fluorescence properties of the dispersions of dimyristoylphosphatidylcholesta-5,7,9-trienol. The phase-transition properties of DRCh are an ideal illustration of calcium-induced isothermal phase transition.

Cholestanes↗
PubMed1984

Phase transition properties of aqueous dispersions of homologues of all-trans 2,3-dipalmitoylcyclopentano-1-phosphocholine.

In a previous publication, (Singer, M.A., Jain, M.K., Sable, H.Z., Pownall, H.H., Mantulin, W.W., Lister, M.D. and Hancock, A.J. (1983) Biochim. Biophys. Acta, 731, 373-377), we reported the properties of aqueous dispersions of the six diastereo-isomers of cyclopentanoid analogues of dipalmitoylphosphatidylcholine. Two of these isomers displayed unusually high enthalpies of transition, about double that of dipalmitoylphosphatidylcholine. One of the high enthalpy isomers whose configuration is all-trans has now been modified by the insertion of extra methylene residues (n = 3 through 9) between the nitrogen and phosphorus atoms of the headgroup. Vesicles were formed from these lipids and studied by 22Na permeability measurements, differential scanning calorimetry, fluorescence polarization, 31P-NMR, and freeze-fracture electron microscopy. Vesicles composed of lipids with n = 2 or 3 exhibit a sharp transition at 46 degrees C or 49 degrees C, respectively, and a high enthalpy with no detectable sub- or pretransitions. Lipids with n greater than 3 exhibit a main transition between 38 and 43 degrees C with enthalpies less than 10 kcal/mol and after prolonged coding (more than 3 days at 4 degrees C) a broad endotherm at about 20 +/- 3 degrees C with enthalpies greater than 4 kcal/mol. These same dispersions display a permeability peak at 20-25 degrees C and a second increase in 22Na efflux in the temperature range 30-40 degrees C. The results of 31P-NMR measurements suggest that the acyl chains in 2,3-dipalmitoylcyclopentano-1-phosphocholine (n = 2) bilayers have restricted rotation below the main phase transition temperature.

Isomerism↗
PubMed1984

Phosphatidylcholine bilayers: subtransitions in pure and in mixed lipids.

Aqueous dispersions of C14, C16, C17 and C18 phosphatidylcholines (PC, where Cn denotes di-acyl of n carbons per chain), and mixtures of C14/C16PC and C16/C17PC were prepared and their thermal properties studied by differential scanning calorimetry (d.s.c.) after sample storage at 2-6 degrees C for up to 22 days. C16PC and C18PC display subtransitions at 22 degrees C and 29 degrees C, respectively, as previously reported by Chen et al. (Proc. Natl. Acad. Sci. U.S.A. 77 (1980) 5060-5063). C17PC shows two subtransitions at 21 degrees C and 26 degrees C, respectively, which are independent of each other. Although C16PC and C17PC individually develop subtransitions, an equimolar mixture does not. However, mixtures of C14/C16PC containing 10 or more mol% of C14PC do display a subtransition. These results underscore the primary dependence of subtransition formation in phosphatidylcholine dispersions on acyl chain structure.

Lipid Bilayers↗
PubMed1983

The properties of membranes formed from cyclopentanoid analogues of phosphatidylcholine.

We have examined the thermal characteristics and barrier properties of vesicles formed from six analogues of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). These analogues differ from DPPC in that the glycerol backbone has been replaced by each of the diastereoisomeric cyclopentane-1,2,3-triols. All of these compounds have main gel to liquid-crystal phase transition temperatures within 5 Kelvin of DPPC and four possess comparable enthalpies and entropies of transition. For two of the analogous, however, the values of the enthalpy and entropy of transition are more than double that of DPPC. The permeability characteristics and organization (as measured by diphenylhexatriene fluorescence depolarization) of vesicles formed from these two compounds suggest that their large transition enthalpy and entropy result from either a reorganization of the polar head group region during the transition or interdigitation of the acyl chains of opposing monolayers.

Cyclopentanes↗
PubMed1983

Interaction of drugs with a model membrane protein. Effects of local anesthetics on electron transfer and hydrogen ion uptake in ionophore stimulated cytochrome oxidase proteoliposomes.

Cytochrome oxidase extracted from beef heart was incorporated into vesicles composed of soy bean phospholipids (asolectin). The oxidation of externally added cytochrome c by such vesicles is associated with proton uptake from the external medium. The rates of both cytochrome c oxidation and proton uptake were stimulated by addition of ionophores such as trifluoromethoxy carbonyl cyanide phenylhydrazone (FCCP), nigericin and valinomycin. These agents probably dissipate pH and/or electrical potential gradients which develop as a result of enzyme activity and which have a "restraining" influence on the turnover of the oxidase. Local anesthetics inhibited oxidase activity but had a much greater effect on the stimulated (ionophore-treated) than the unstimulated enzyme. In addition, pretreating proteoliposomes with local anesthetics completely prevented the stimulating effects of these ionophores. Based on this and previous studies, a model was developed in which local anesthetics interacted with the phospholipid component of the oxidase complex resulting in reduced internal electron transfer and dissociation of the oxidase from the regulatory role of the proton gradient.

Anesthetics, Local↗
PubMed1982

Interaction of drugs with a model membrane protein. Effect of dibucaine on cytochrome oxidase proteoliposomes.

Cytochrome oxidase is a mitochondrial trans-membrane protein which catalyzes the vectorial transfer of electrons from cytochrome c to molecular oxygen. When the oxidase was incorporated into liposomes composed of saturated phospholipids, enzymatic activity was reduced as compared to the activity of either the isolated enzyme or the enzyme incorporated into soy bean phospholipid (asolectin) liposomes. This reduced activity probably resulted from partial replacement of retained oxidase boundary lipid with exogenously added lipid and an unfavourable orientation of a portion of the oxidase molecules for reaction with externally added substrate. On the other hand, substrate binding at the low affinity site was enhanced by incorporation of the oxidase into vesicles composed of either saturated phospholipids or asolectin. At pH 7.4 the local anesthetic dibucaine behaved as an uncompetitive inhibitor of the enzyme, while at pH 6.0 the inhibition pattern became mixed in type. Dibucaine had similar effects on both the isolated and incorporated enzyme except that, in general, the anesthetic caused less inhibition of the incorporated oxidase. It is postulated that positively charged anesthetic molecules act predominantly by competing with substrate for binding while non-charged anesthetic molecules interact with the oxidase boundary lipid to form non-productive complexes.

Animals↗
PubMed1982

Investigation of the membrane-fluidizing properties of porphyrin-inducing drugs.

The objective of this study was to test the hypothesis that porphyrin-inducing drugs act at least in part by disrupting membrane lipids. The porphyrin-inducing steroids, 3 alpha-hydroxy-5 alpha-pregnane-11,20-dione (alfaxalone) and 3 alpha-hydroxy-5 alpha-pregnan-20-one induce considerably greater fluidity changes in spin-labelled phospholipid-cholesterol bilayers than do the corresponding 3 beta-hydroxy steroids which are also less potent as porphyrin inducers. The steroids did not cause any significant change in spin-labelled vesicles lacking cholesterol. The porphyrin-inducing compound 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine and a series of analogues caused fluidity changes in phospholipid bilayers in the presence and absence of cholesterol. The porphyrin-inducing nonplanar 2,2',4,4'-6,6'-hexachlorobiphenyl caused a significant change in bilayer fluidity in phospholipid bilayers in the presence and absence of cholesterol. Since the planar 3,3',4,4'-tetrachlorobiphenyl, allylisopropylacetamide (AIA), and griseofulvin are potent porphyrin-inducing compounds, but do not fluidize a lipid bilayer, it was clear that the original hypothesis required modification. No evidence could be obtained to support the idea that a subset of porphyrin-inducing drugs exists which are membrane fluidizers and whose common mechanism of action as porphyrin-inducers might be revealed by a common pattern of porphyrin accumulation in chick embryo liver cells. It is suggested that those porphyrin-inducing compounds with membrane-fluidizing properties might fluidize the nuclear membrane, thus facilitating the transfer of an induction specific RNA for delta-aminolevulinic acid synthetase from the nucleus to the cytoplasm.

Allylisopropylacetamide↗
PubMed1980

Interaction of four local anesthetics with phospholipid bilayer membranes: permeability effects and possible mechanisms.

Sodium 22 efflux was measured, at various temperatures, in the presence of different local anesthetics (dibucaine, tetracaine, benzocaine, procaine) in multilamellar liposomes composed of saturated phosphatidylcholines and dicetyl phosphate. These liposomes display a permeability maximum in the temperature region of their respective phase transitions. Local anesthetics cause this permeability maximum to occur at a lower temperature and also increase its magnitude. Binding of the anesthetics to these liposomes displays a biphasic temperature curve with a maximum in the vicinity of the transition temperature. In addition there appears to be a linear relationship between aqueous anesthetic concentrations causing equal permeability effects and corresponding membrane concentrations. All of the anesthetics decrease the cooperativity of the lipid phase transition and this is probably an important factor underlying induced permeability effects. However several observations suggest that additional factors are also important and we have postulated that one such factor is the preferential binding of the anesthetics to phase boundary regions.

Anesthetics, Local↗
PubMed1980

Use of computerized tomography to evaluate bleeding after renal biopsy.

Perirenal bleeding following biopsy was assessed in 25 consecutive cases using computerized axial tomography (CT) scanning. Perirenal hematomas were found in 15 patients (60%). In 8 patients the hematomas were moderate or large in size. There was no correlation between the clinical findings, fall in hemoglobin, presence of macroscopic hematuria and the finding of perirenal hematoma by CT scanning.

Adolescent↗