Effect of Inhalation Anesthetics on Spin-Labeled Cholesterol Containing DPPC Vesicles

1988 ◽  
Vol 43 (3-4) ◽  
pp. 264-268 ◽  
Author(s):  
N. Gulfo ◽  
R. Bartucci ◽  
L. Sportelli

We have investigated by means of electron spin resonance (ESR) spectroscopy the influence of three inhalation anesthetics, i.e. halothane, chloroform and diethyl ether, on the interfacial and hydrophobic region as well of 38 mol% cholesterol containing DPPC unilamellar vesicles. The study has been carried out in the temperature range 25-45 °C. The variation of the order parameter, S, vs temperature of the lipid phase indicates that with this content of cholesterol the characteristic gel → liquid crystalline main phase transition of DPPC, normally occurring at Tt ~ 41 °C, disappears. When halothane and chloroform are added to the vesicles suspension up to [DPPC]/[anesthetic] molar ratio of 1:1 the main phase transition, as detected with the stearic acid spin label I(12,3), reappears again and it results down shifted at Tt ~ 35 and 39 °C, respectively. In presence of diethyl ether, instead, the main phase transition is not observable also at the highest concentration of anesthetic used. Moreover, halothane and chloroform affect similarly the hydrophobic core of cholesterol-!- DPPC vesicles which, in turn, results to be different from the action exerted by diethyl ether in the same region. The ESR findings are discussed in terms of competitive effects shown by cholesterol and inhalation anesthetics. Moreover, the interfacial region of CHOL + DPPC vesicles results to be the target of anesthetics.

2018 ◽  
Vol 115 (32) ◽  
pp. E7587-E7594 ◽  
Author(s):  
Nicolò Paracini ◽  
Luke A. Clifton ◽  
Maximilian W. A. Skoda ◽  
Jeremy H. Lakey

The outer membrane (OM) of Gram-negative bacteria is a robust, impermeable, asymmetric bilayer of outer lipopolysaccharides (LPSs) and inner phospholipids containing selective pore proteins which confer on it the properties of a molecular sieve. This structure severely limits the variety of antibiotic molecules effective against Gram-negative pathogens and, as antibiotic resistance has increased, so has the need to solve the OM permeability problem. Polymyxin B (PmB) represents those rare antibiotics which act directly on the OM and which offer a distinct starting point for new antibiotic development. Here we investigate PmB’s interactions with in vitro OM models and show how the physical state of the lipid matrix of the OM is a critical factor in regulating the interaction with the antimicrobial peptide. Using neutron reflectometry and infrared spectroscopy, we reveal the structural and chemical changes induced by PmB on OM models of increasing complexity. In particular, only a tightly packed model reproduced the temperature-controlled disruption of the asymmetric lipid bilayer by PmB observed in vivo. By measuring the order of outer-leaflet LPS and inner-leaflet phospholipids, we show that PmB insertion is dependent on the phase transition of LPS from the gel to the liquid crystalline state. The demonstration of a lipid phase transition in the physiological temperature range also supports the hypothesis that bacteria grown at different temperatures adapt their LPS structures to maintain a homeoviscous OM.


Molecules ◽  
2021 ◽  
Vol 26 (10) ◽  
pp. 2851
Author(s):  
Grażyna Neunert ◽  
Jolanta Tomaszewska-Gras ◽  
Aneta Baj ◽  
Marlena Gauza-Włodarczyk ◽  
Stanislaw Witkowski ◽  
...  

Steady-state emission spectroscopy of 1-anilino-8- naphthalene sulfonate (ANS) and 1,6-diphenyl-1,3,5-hexatriene (DPH), fluorescence anisotropy, and DSC methods were used to characterize the interactions of the newly synthesized 1-carba-alpha-tocopherol (CT) with a 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) membrane. The DSC results showed significant perturbations in the DPPC structure for CT concentrations as low as 2 mol%. The main phase transition peak was broadened and shifted to lower temperatures in a concentration-dependent manner, and pretransition was abolished. Increasing CT concentrations induced the formation of new phases in the DPPC structure, leading to melting at lower temperatures and, finally, disruption of the ordered DPPC structure. Hydration and structural changes of the DPPC liposomes using ANS and DPH fluorescent probes, which are selectively located at different places in the bilayer, were studied. With the increased concentration of CT molecules in the DPPC liposomes, structural changes with the simultaneous formation of different phases of such mixture were observed. Temperature studies of such mixtures revealed a decrease in the temperature of the main phase transition and fluidization at decreasing temperatures related to increasing hydration in the bilayer. Contour plots obtained from concentration–temperature data with fluorescent probes allowed for identification of different phases, such as gel, ordered liquid, disordered liquid, and liquid crystalline phases. The CT molecule with a modified chromanol ring embedded in the bilayer led to H-bonding interactions, expelling water molecules from the interphase, thus introducing disorder and structural changes to the highly ordered gel phase.


1990 ◽  
Vol 68 (1) ◽  
pp. 161-168 ◽  
Author(s):  
Rajan George ◽  
Ruthven N.A.H. Lewis ◽  
Ronald N. McElhaney

The purified Na+, Mg2+-ATPase from the Acholeplasma laidlawii B plasma membrane was reconstituted with dimyristoyl phosphatidylcholine and the lipid thermotropic phase behavior of the proteoliposomes formed was investigated by differential scanning calorimetry. The effect of this ATPase on the host lipid phase transition is markedly dependent on the amount of protein incorporated. At low protein/lipid ratios, the presence of increasing quantities of ATPase in the proteoliposomes increases the temperature and enthalpy while decreasing the cooperativity of the dimyristoyl phosphatidylcholine gel to liquid–crystalline phase transition. At higher protein/lipid ratios, the incorporation of increasing amounts of this enzyme does not further alter the temperature and cooperativity of the phospholipid chain-melting transition, but progressively and markedly decreases the transition enthalpy. Plots of lipid phase transition enthalpy versus protein concentration suggest that at the higher protein/lipid ratios each ATPase molecule removes approximately 1000 dimyristoyl phosphatidylcholine molecules from participation in the cooperative gel to liquid–crystalline phase transition of the bulk lipid phase. These results indicate that this integral transmembrane protein interacts in a complex, concentration-dependent manner with its host phospholipid and that such interactions involve both hydrophobic interactions with the lipid bilayer core and electrostatic interactions with the lipid polar head groups at the bilayer surface.Key words: Acholeplasma laidlawii B, Na+,Mg2+-ATPase, differential scanning calorimetry, lipid-protein interactions.


2006 ◽  
Vol 73 (2) ◽  
pp. 415-420 ◽  
Author(s):  
Patricia Castellano ◽  
Graciela Vignolo ◽  
Ricardo Norberto Farías ◽  
José Luis Arrondo ◽  
Rosana Chehín

ABSTRACT Lactocin 705 is a bacteriocin whose activity depends upon the complementation of two peptides, termed Lac705α and Lac705β. Neither Lac705α nor Lac705β displayed bacteriocin activity by itself when the growth of sensitive cells was monitored. To obtain molecular insights into the lactocin 705 mechanism of action, Fourier transform infrared spectroscopy was used to investigate the interactions of each peptide (Lac705α and Lac705β) with dipalmitoylphosphatidylcholine liposomal membranes. Both peptides show the ability to interact with the zwitterionic membrane but at different bilayer levels. While Lac705α interacts with the interfacial region inducing dehydration, Lac705β peptide interacts with only the hydrophobic core. This paper presents the first experimental evidence that supports the hypothesis that Lac705α and Lac705β peptides could form a transmembrane oligomer. From the obtained results, a mechanism of action of lactocin 705 on membrane systems is proposed. The component Lac705α could induce the dehydration of the bilayer interfacial region, and the Lac705β peptide could insert in the hydrophobic region of the membrane where the peptide has adequate conditions to achieve the oligomerization.


1979 ◽  
Vol 34 (5-6) ◽  
pp. 406-413 ◽  
Author(s):  
J. Luisetti ◽  
H. Möhwald ◽  
H. J. Galla

Abstract Absorption and fluorescence experiments on pheophytin and chlorophyll containing lipid bilayer vesicles are reported. Pheophytin aggregates on the vesicles are established from an additional red shifted band (at 695 nm) in the absorption spectrum. These aggregates contain pheophytin in an arrangement with the molecular planes of the porphyrin rings being parallel and cover about 10% of the vesicle surface. The lipid phase dissolves pheophytin up to a molar ratio of 15% above the lipid phase transition. This solubility limit decreases hardly on solidification of the lipid.For chlorophyll a containing vesicles the aggregates are not observed in the absorption spectrum. The chlorophyll solubility is about equal to that of pheophytin. This suggests that the phase separa­ tion indicated from fluorescence measurements at temperatures below the lipid phase transition does not lead to the formation of strongly bound chlorophyll aggregates.


2013 ◽  
Vol 652-654 ◽  
pp. 459-462
Author(s):  
Ya Tong Guo ◽  
Zhu Zheng ◽  
Zhen Qi Hou ◽  
Jie Du

A series of biodegradable segmented liquid crystalline poly(ester-urethane)s were prepared by solution polymerization of poly(L-lactic acid) (PLLA), mesogenic diol prepolymer poly(butylene terephthaloyldioxy dibenzoates) (MD), and hexamethylene diisocyanate (HDI). The MD content was varied from 0 to 40 mol% so that the effects of the mesogen content on the thermal and physical properties, and hydrolytic degradation were examined respectively. It was found that introducing mesogens units could increase the thermal stability and the elastic properties, while reduced the phase transition temperatures and the hydrolytic degradation rate.


1984 ◽  
Vol 62 (11) ◽  
pp. 1134-1150 ◽  
Author(s):  
P. M. Macdonald ◽  
B. D. Sykes ◽  
R. N. McElhaney

The orientational order parameters of monofluoropalmitic acids biosynthetically incorporated into membranes of Acholeplasma laidlawii B in the presence of a large excess of a variety of structurally diverse fatty acids have been determined via 19F nuclear magnetic resonance (19F NMR) spectroscopy. It is demonstrated that these monofluoropalmitic acids are relatively nonperturbing membrane probes based upon physical (differential scanning calorimetry), biochemical (membrane lipid analysis), and biological (growth studies) criteria. 19F NMR is shown to convey the same qualitative and quantitative picture of membrane lipid order provided by 2H-NMR techniques and to be sensitive to the structural characteristics of the membrane fatty acyl chains, as well as to the lipid phase transition. Representatives of each naturally occurring class of fatty acyl chain structures, including straight-chain saturated, methyl-branched, monounsaturated, and alicyclic-ring-substituted fatty acids, were studied and the 19F-NMR order parameters were correlated with the lipid phase transitions (determined calorimetrically). The lipid phase transition was the prime determinant of overall orientational order regardless of fatty acid structure. Effects upon orientational order attributable to specific structural substituents were discernible, but were secondary to the effects of the lipid phase transition. In the gel state, relative overall order was directly proportional to the temperature of the particular lipid phase transition. Not only the overall order, but also the order profile across the membrane was sensitive to the presence of particular structural substituents. In particular, in the gel state specific fatty acyl structures demonstrated a characteristic disordering effect in the membrane order profile. These various observations can be merged to provide a unified picture of the manner in which fatty acyl chain chemistry modulates the physical state of membrane lipids.


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