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Biomedical subjects

I R Miller

Publications and source records attributed to I R Miller.

At least 19 recordsLinked to original sources
PubMed2005

Hydration of phospholipid bilayers in the presence and absence of cholesterol.

The number of water molecules bound (unfreezable) by a molecule of dipalmitoyl phosphatidylserine (DPPS) or by a molecule of dipalmitoyl phosphatidylcholine (DPPC) alone or in mixtures with cholesterol was determined by differential scanning calorimetry (DSC). When the phospholipids are in the gel state and in the absence of cholesterol, molecule of DPPS binds about 3.5 molecules of water and molecule of DPPC binds about 6 molecules of water. Number of water molecules bound increases when cholesterol crystallites are formed in the bilayer. For DPPS-cholesterol mixture at X(chol) -0.5, as well as for DPPC-cholesterol mixture at X(chol) -0.5 about 7 water molecules are bound.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2001

Attenuated total reflection (ATR) Fourier transform infrared spectroscopy of dimyristoyl phosphatidylserine-cholesterol mixtures.

Mixtures of cholesterol with dimyristoyl phosphatidylserine or deuterated dimyristoyl phosphatidylserine were investigated by polarized and non polarized attenuated total reflection (ATR) Fourier transform infrared (FTIR) Spectroscopy. From polarized spectra the dichroic ratios of various vibrations as a function of cholesterol were calculated. Dichroic ratios of methylene vibration (CH(2)) 2934 cm(-1) of cholesterol decreases with increase of cholesterol concentration leveling off in the region where cholesterol phase separation takes place. The orientation of deuterated methylene (CD(2)) symmetric and asymmetric bands of the deuterated dimyristoyl phosphatidylserine is influenced little by cholesterol. In the polar region of dimyristoyl phosphatidylserine no effect of cholesterol on the dichroic ratios of carbonyl (C==O) and asymmetric phosphate (PO(2)(-)) vibrations were detected. For nonpolarized spectra the broad bands in the polar region of the phospholipid were deconvoluted. The carbonyl band (C==O) in pure dimyristoyl phosphatidylserine is composed of five bands; in the presence of increasing concentrations of cholesterol conformational change of these vibrations takes place evolving into one predominant band. Similar conformational change takes place in the presence of 75 molecules water/molecule DMPS. For the asymmetric phosphate band very small shifts due to interaction with cholesterol were detected.

Cholesterol↗
PubMed2000

Organization of water molecules by adhering to oriented layers of dipalmitoylphosphatidyl serine in the presence of varying concentrations of cholesterol.

About seven water molecules adhere to one molecule of dipalmitoylphosphatidyl serine (DPPS) in an oriented surface layer as inferred from the increase of the dichroic ratio R of their OH stretching vibration band (3400 cm(-1)) from 2 in the random bulk state to about 2.8 when adhering to DPPS. In DPPS-cholesterol mixtures the number of water molecules adhering to the phospholipid molecules and oriented by them increases as cholesterol content increases. This increase is very steep between molar fractions of cholesterol X(chol)=0.2-0.4 and at X(chol)=0.6 about 13 water molecules adhere and are oriented by one DPPS molecule.

Cholesterol↗
PubMed2000

Structure of phosphatidyl serine (PS) involved in interbilayer salt bridging and hydrogen bonding.

For understanding the experimental results indicating salt bridging and hydrogen bonding between opposite polar surfaces in planar multibilayer structures of phosphatidyl serine (PS), (1) we carried out molecular modeling of the interacting surface layers. The interacting structures in the planar multibilayers are stabilized by salt bridge arrays of the phosphates with their counterions and by hydrogen bonds of ammonia from one polar surface with carbonyls in the opposite one. In multishell liposomes, where the distances between phosphates on the facing each other surfaces are not equal and they are bound to get out of register, the interbilayer interaction cannot extend over a large enough area to form stable structures, except if the salt bridges are strong enough to break down the curved surfaces and form planar multibilayers.

Hydrogen Bonding↗
PubMed1999

Hydration of phosphatidyl serine multilayers and its modulation by conformational change induced by correlated electrostatic interaction.

Hydration of the various residues of phospholipids was inferred from the shift in the wave number of their vibration bands, obtained from the amplitudes of their positive and negative peaks in the difference spectra between those of the hydrated and the dry phospholipid multibilayers. The effect of aligned phospholipid layers on the orientation of their hydrating water molecules was inferred from the dichroic ratio of the OH stretching band, measured by polarized attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) with a germanium prism, as a function of the water-to-lipid ratio in the surface film. The results indicate that about seven water molecules are oriented by one phosphatidyl serine molecule in the surface film. About 8 to 11 additional water molecules contribute to the hydration of the polar residues as revealed by the effect on the difference spectra. The hydration appears to be cooperative. A water molecule that initiates hydration of a site facilitates access of additional water molecules, until the hydration of the whole site composed of many different interacting polar residues is completed.

Lipid Bilayers↗
PubMed1998

Hydration of phospholipid bilayers in the presence and absence of cholesterol.

Differential scanning calorimetry (DSC) was used for determining the number of unfreezable water molecules per molecule of phosphatidylserine from bovine spinal cord (PS) or dimyristoyl phosphatidylserine (DMPS) and dimyristoyl phosphatidylcholine (DMPC), alone or in mixtures with cholesterol. It was assumed that the unfreezable water molecules are tightly bound to the phospholipid. It was found that when the phospholipids are in the gel state and in the absence of cholesterol, PS binds 2.5 water molecules, DMPS 3.8 water molecules and DMPC 3.5 water molecules. In the presence of cholesterol the number of water molecules bound increases in the region where phase separation of cholesterol takes place [D. Bach, Chem. Phys. Lipids 35 (1984) 385-392; E.J. Wachtel, N. Borochov, D. Bach, Biochim. Biophys. Acta 1066 (1991) 63-69; D. Bach, N. Borochov, E. Wacktel, Chem. Phys. Lipids, submitted].

Animals↗
PubMed1998

The effect of ethanol on the structure of phosphatidylserine bilayers.

Thermotrophic and structural effects of ethanol on phosphatidylserine (PS) membranes were investigated by differential scanning calorimetry (DSC) and X-ray diffraction. It was found that up to 15% (v/v) added ethanol, there is little change in the melting temperature of the phospholipid and no change in the interbilayer (d) spacing in the gel phase, indicating that there is no interdigitation of the hydrocarbon chains. Above the melting temperature of the phospholipid, a large decrease of the d spacing, due primarily to a decrease in the thickness of the bilayer, was found. Ethanol molecules located in the headgroup region apparently expand the area available to the headgroups with concomitant coiling of the acyl chains, resulting in marked thinning of the lipid layer.

Anesthetics, General↗
PubMed1996

Structure and orientation of the mammalian antibacterial peptide cecropin P1 within phospholipid membranes.

Cecropins are positively charged antibacterial peptides that act by permeating the membrane of susceptible bacteria. To gain insight into the mechanism of membrane permeation, the secondary structure and the orientation within phospholipid membranes of the mammalian cecropin P1 (CecP) was studied using attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy and molecular dynamics simulations. The shape and frequency of the amide I and II absorption peaks of CecP within acidic PE/PG multibilayers (phosphatidylethanolamine/phosphatidylglycerol) in a 7:3 (w/w) ratio (a phospholipid composition similar to that of many bacterial membranes), indicated that the peptide is predominantly alpha-helical. Polarized ATR-FTIR spectroscopy was used to determine the orientation of the peptide relative to the bilayer normal of phospholipid multibilayers. The ATR dichroic ratio of the amide I band of CecP peptide reconstituted into oriented PE/PG phospholipid membranes indicated that the peptide is preferentially oriented nearly parallel to the surface of the lipid membranes. A similar secondary structure and orientation were found when zwitterionic phosphatidylcholine phospholipids were used. The incorporation of CecP did not significantly change the order parameters of the acyl chains of the multibilayer, further suggesting that CecP does not penetrate the hydrocarbon core of the membranes. Molecular dynamics simulations were used to gain insight into possible effects of transmembrane potential on the orientation of CecP relative to the membrane. The simulations appear to confirm that CecP adopts an orientation parallel to the membrane surface and does not insert into the bilayer in response to a cis positive transmembrane voltage difference. Taken together, the results further support a "carpet-like" mechanism, rather than the formation of transmembrane pores, as the mode of action of CecP. According to this model, formation of a layer of peptide monomers on the membrane surface destablizes the phospholipid packing of the membrane leading to its eventual disintegration.

Amino Acid Sequence↗
PubMed1993

Thermotropic behavior of phosphatidylcholine-glucosyl ceramide mixtures: effects of phospholipid acyl chain composition and interaction with water.

The thermotropic behavior of multilamellar vesicles composed of mixtures of dimyristoyl phosphatidylcholine-glucosyl ceramide and of egg phosphatidylcholine-glucosyl ceramide was investigated using differential scanning calorimetry. Macroscopic demixing of the lipid components occurred when multilamellar vesicles were prepared from mixtures of glucosyl ceramide and egg phosphatidylcholine by conventional methods. This problem was overcome by a technique based on spray drying of the lipid mixture. The results obtained for the two systems are compared with data available for dipalmitoyl phosphatidylcholine-glucosyl ceramide mixtures (Biochemistry 22 (1983) 3497-3501). All three phosphatidylcholines perturb the complex thermotropic behavior of glucosyl ceramide. The data suggest that the interference with intermolecular interactions among glycosyl ceramide molecules by phospholipid molecules is related to the molecular miscibility of the two components. This is strongly dependent on the acyl chain composition of the phosphatidylcholine and the water activity of the ambient aqueous phase.

Calorimetry, Differential Scanning↗
PubMed1991

Fourier transform infrared spectroscopy of aqueous dispersions of phosphatidylserine-cholesterol mixtures.

The effect of cholesterol on vibrational spectra in the non polar and in the polar region of dimyristoyl phosphatidylserine (DMPS) and of phosphatidylserine from bovine spinal cord (PS) has been investigated. The small shifts in the methylene CH stretching frequencies after taking into account the contribution of the cholesterol spectrum were interpreted as a combined effect of cholesterol on the conformation of the chains and of the lesser contributions of the cholesterol methyl groups. Cholesterol also influences the ratio of the trans (1465 cm-1) to the lower wavelength (1457 cm-1) CH2 bending bands. No significant direct effect of cholesterol on the vibration of the polar residues was discerned. The small shift of the carboxylate band observed below the phase transition is probably due to the change in the intermolecular zwitterions when the average distance between the neighboring polar groups increases due to incorporation of cholesterol molecules.

Animals↗
PubMed1990

Lipid dependence of surface conformations of protein kinase C.

The change of conformation of protein kinase C interacting with the surface of a mercury electrode directly from a solution or through a lipid monolayer was inferred from the number of cystine residues exposed and reduced on the electrode and from their reduction potentials. Soluble protein kinase C was estimated to have 5-6 disulfide bonds which could potentially react with the mercury electrode. Two major reduction peaks of cystine at different microenvironments within the protein molecule adsorbed to a mercury surface. They were observed in a.c. polarograms and cyclic voltamograms at two distinct potentials. The potential of these peaks became more negative as the pH of the solution increased, which was consistent with relaxation or decrease in alpha-helicity (ordered structure) of the protein as determined by circular dichroism (CD) estimations of secondary structure. The peak at the more positive potentials (-0.46 V relative to NAg/AgCl electrode at pH 7.4) tended to vanish upon cyclic reduction and reoxidation of the cystine, while the more negative peak (-0.62 V at pH 7.4) was enhanced. Addition of Mg2+ or Ca2+ had no significant effect on the potential but there was a reduction in their amplitude which appeared to affect the disappearance of these peaks upon pH adjustment. This suggests that the tertiary structure of the molecule is stabilized by Ca2+ and Mg2+, as substantiated by CD spectral analysis of secondary structures. Protein kinase C penetrated lipid monolayers to some extent. Addition of diacylglycerol or phorbol ester to the lipid monolayers facilitated this penetration. These compounds stabilized the protein surface conformation by destabilizing the monolayer at more positive potentials, resulting in an enhanced reduction peak at -0.42 V. This phenomenon was not significantly affected by Mg2+ or by Ca2+. The region of the protein kinase C (PKC) sequence which penetrated the monolayer contains cysteines and a primary amine(s), and may have homology to a region of phospholipase A2 which has been proposed as a phospholipid binding site for the two enzymes. Additionally, these polarographic studies suggest that PKC associates with and penetrates monolayers in a divalent cation-independent manner in agreement with our previous physical analyses of PKC interactions with lipid bilayers.

Adsorption↗
PubMed1990

Electric field dependence of alamethicin channels.

Circular dichroism (CD) of alamethicin embedded in vesicular membranes from outside, and its change, upon imposing Donnan potentials across the membrane, was measured. The changes in CD suggested a decrease in a helicity and increase in beta structure with the membrane potential positive inside and vice versa when the potential was positive on the outer side of the vesicles from where the alamethicin was inserted into the membrane. The Donnan potential was created by entrapping the polyacrylate (PA-) in the vesicles and changing the salt concentration outside or by adding different concentrations of PA- or polyethyleneimide (PEI+) at the outside of vesicles with 2 x 10(-5) M salt inside. The effect of the potential on the CD spectra and thus the alamethicin conformation is independent on the type of the polyelectrolyte employed for the Donnan potential generation.

Alamethicin↗
PubMed1988

Interaction of furosemide with lipid membranes.

The interaction of furosemide with different phospholipids was investigated. Its influence on the lipid structure was inferred from its effect on the phase transition properties of lipids and on the conductance of planar bilayer membranes. The thermotropic properties of dipalmitoyl phosphatidylcholine, phosphatidylethanolamine (natural), dipalmitoyl phosphatidylethanolamine, brain sphingomyelin, brain cerebrosides and phosphatidylserine in the presence and absence of furosemide were investigated by differential scanning calorimetry. The modifying effect of furosemide seems to be strongest on phosphatidylethanolamine (natural) and sphingomyelin bilayers. The propensity of furosemide to decrease the electrical resistance of planar lipid membranes was also studied and it is shown that the drug facilitates the transport of ions. Partition coefficients of furosemide between lipid bilayers and water were measured.

Cerebrosides↗
PubMed1988

Penetration of furosemide into phospholipid monolayers.

Furosemide is a surface-active anion and it tends to displace lipid monolayers from the surface at positive polarizations lowering their potential stability range. The efficiency of the penetration and the displacement increases with decreasing surface pressure of the monolayer. Lower capacitance at a wider potential range corresponds to higher surface pressure. Monolayers with higher capacitances are indeed more readily penetrated and displaced as demonstrated by further increase in their capacitance and increase in their proton conductance. Furosemide raises the capacitance of the monolayer in the stable region due to intercalation between the head groups thus reducing the thickness of the hydrocarbon layer. In pure PC monolayer about 10% increase in capacitance is observed in the presence of 6 X 10(-4)M furosemide. The effect of furosemide becomes more pronounced with increasing sphingomyelin content in the mixed monolayers. The monolayer of PE is more condensed and its capacitance is lower (approximately 1.45 microF/cm2) and is stable in a wider potential range than that of PC. It is less affected by furosemide and concentrations higher than 10(-3) M are required to narrow the stability range and to increase the capacitance.

Furosemide↗
PubMed1988

X-ray-induced specific locus mutations in the ad-3 region of two-component heterokaryons of Neurospora crassa. I. Modification of the heterozygous effects of multilocus deletions covering the ad-3A or ad-3B loci.

The basis for the reduced growth rates of heterokaryons between strains carrying nonallelic combinations of gene/point mutations (ad-3R) and multilocus deletion mutations (ad-3IR) has been investigated by a simple genetic test. The growth rates of forced 2-component heterokaryons (dikaryons) between multilocus deletion mutations were compared with forced 3-component heterokaryons (trikaryons) containing an ad-3AR ad-3BR double mutant as their third component. Since the third component has no genetic damage at other loci immediately adjacent to the ad-3A or ad-3B locus, the growth rate on minimal medium depends on the functional activity of the unaltered (and presumed "wild-type") ad-3A and ad-3B loci in the first two components. In many cases, the requirements of the original dikaryons have been satisfied by the addition of unaltered genes (in the third component), and these trikaryons grow at wild-type rate on minimal medium. Those trikaryons growing at less than wild-type rate were shown to be adenine-requiring, and wild-type growth rate was obtained with the addition of low levels of adenine to the medium. Such tests in the present experiments have shown that ad-3IR mutations result not only in inactivation of the ad-3 loci by multilocus deletion but also, in many cases, in partial gene inactivation by an unknown mechanisms at other loci in the immediately adjacent regions. The heterozygous effects observed in our present experiments with multilocus deletions in Neurospora can be explained either by a spreading-type position effect of the type found by others in Drosophila, mice, Oenothera and Aspergillus or by undetected genetic damage ("cryptic mutations") in the immediately adjacent genetic regions. An attempt will be made to distinguish between these two alternative hypotheses with techniques for DNA cloning and sequencing in future experiments.

Adenine↗
PubMed1987

Zwitterionic water chains as H+/OH- transporters.

Zwitterionic water chains spanning the hydrocarbon layer with an H+ attached to one end and an OH- to the other end are suggested to be H+ and OH- conductors. They explain higher H+/OH- fluxes and their very weak dependence on pH.

Hydrogen Bonding↗
PubMed1986

Effect of ethylene glycol on the phase transition kinetics of gluco- and galactocerebrosides.

The effect of different concentrations of ethylene glycol in water on the phase transition (metastable----stable state) of Gaucher's glucocerebroside, of bovine brain cerebroside type II (non hydroxy acyl chains only) and of N-palmitoylgalactocerebroside has been investigated. The phase transition and its kinetics were inferred from the thermograms at different heating and cooling rates and confirmed by FTIR spectra of the cerebrosides in the different states. The significance of the conformational differences of the glucose and of the galactose residues with respect to their solvation, and the subsequent effect on the intermolecular interactions and the phase transition is discussed.

Calorimetry, Differential Scanning↗